U.S. patent number 10,342,794 [Application Number 15/945,685] was granted by the patent office on 2019-07-09 for substituted imidazo[1,2-a]pyrazines as syk inhibitors.
This patent grant is currently assigned to Gilead Sciences, Inc.. The grantee listed for this patent is Gilead Sciences, Inc.. Invention is credited to Peter A. Blomgren, Kevin S. Currie, Jeffrey E. Kropf, Seung H. Lee, Jennifer R. Lo, Scott A. Mitchell, Aaron C. Schmitt, Sundaramoorthi Swaminathan, Jin-Ming Xiong, Jianjun Xu, Zhongdong Zhao.
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United States Patent |
10,342,794 |
Blomgren , et al. |
July 9, 2019 |
Substituted imidazo[1,2-a]pyrazines as Syk inhibitors
Abstract
The present disclosure relates to compounds that are Syk
inhibitors and to their use in the treatment of various disease
states, including cancer and inflammatory conditions. In particular
embodiments, the structure of the compounds is given by Formula I:
##STR00001## wherein R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are as
described herein. The present disclosure further provides
pharmaceutical compositions that include a compound of Formula I,
or pharmaceutically acceptable salts or co-crystals thereof, and
methods of using these compounds and compositions to treat
conditions mediated by Syk.
Inventors: |
Blomgren; Peter A. (Issaquah,
WA), Currie; Kevin S. (North Bend, WA), Kropf; Jeffrey
E. (Issaquah, WA), Lee; Seung H. (Branford, CT), Lo;
Jennifer R. (Branford, CT), Mitchell; Scott A. (East
Haven, CT), Schmitt; Aaron C. (Hamden, CT), Swaminathan;
Sundaramoorthi (Burlingame, CA), Xiong; Jin-Ming
(Guilford, CT), Xu; Jianjun (Seattle, WA), Zhao;
Zhongdong (Guilford, CT) |
Applicant: |
Name |
City |
State |
Country |
Type |
Gilead Sciences, Inc. |
Foster City |
CA |
US |
|
|
Assignee: |
Gilead Sciences, Inc. (Foster
City, CA)
|
Family
ID: |
52350364 |
Appl.
No.: |
15/945,685 |
Filed: |
April 4, 2018 |
Prior Publication Data
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Document
Identifier |
Publication Date |
|
US 20180325896 A1 |
Nov 15, 2018 |
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Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
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15017394 |
Feb 5, 2016 |
9968601 |
|
|
|
14578973 |
Dec 22, 2014 |
9290505 |
|
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61920407 |
Dec 23, 2013 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P
1/00 (20180101); A61P 7/06 (20180101); A61P
1/16 (20180101); A61P 5/14 (20180101); A61P
31/04 (20180101); A61P 37/08 (20180101); A61P
3/10 (20180101); A61K 9/0053 (20130101); A61P
7/00 (20180101); C07D 487/04 (20130101); A61P
11/06 (20180101); A61P 25/28 (20180101); A61K
31/5377 (20130101); A61P 1/18 (20180101); A61P
11/00 (20180101); A61P 17/04 (20180101); A61P
1/02 (20180101); A61P 19/08 (20180101); A61P
17/00 (20180101); A61P 19/06 (20180101); A61P
37/00 (20180101); A61P 21/02 (20180101); A61P
35/02 (20180101); A61P 13/02 (20180101); A61P
21/04 (20180101); A61P 37/06 (20180101); A61P
37/02 (20180101); A61P 17/06 (20180101); A61P
11/02 (20180101); A61P 21/00 (20180101); A61P
25/16 (20180101); A61P 1/04 (20180101); A61P
9/00 (20180101); A61P 19/00 (20180101); A61P
19/02 (20180101); A61P 35/00 (20180101); A61P
43/00 (20180101); A61P 13/12 (20180101); A61P
17/02 (20180101); A61P 25/00 (20180101); A61P
9/10 (20180101); A61P 27/02 (20180101); A61P
35/04 (20180101); A61P 7/04 (20180101); A61P
29/00 (20180101); A61K 31/4985 (20130101); A61P
11/08 (20180101) |
Current International
Class: |
A61K
31/4985 (20060101); A61K 9/00 (20060101); C07D
487/04 (20060101); A61K 31/5377 (20060101) |
Field of
Search: |
;514/249
;544/117,350,359,405 ;549/510 |
References Cited
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|
Primary Examiner: Willis; Douglas M
Attorney, Agent or Firm: Sheppard Mullin Richter &
Hampton LLP
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of U.S. application Ser. No.
15/017,394, filed Feb. 5, 2016, now U.S. Pat. No. 9,968,601, which
is a continuation of U.S. application Ser. No. 14/578,973, now U.S.
Pat. No. 9,290,505, which claims benefit under 35 U.S.C. .sctn.
119(e) to U.S. Provisional Application No. 61/920,407, filed Dec.
23, 2013, each of which are incorporated herein by reference.
Claims
What is claimed:
1. A crystalline Form I of a monomesylate salt of Compound 2:
##STR00149## wherein the crystalline form is characterized by an
X-ray powder diffractogram comprising peaks (.degree. 2.theta.) at
17.3.degree..+-.0.2.degree. 2.theta., 17.9.degree..+-.0.2.degree.
2.theta., 19.7.degree..+-.0.2.degree. 2.theta.,
21.6.degree..+-.0.2.degree. 2.theta. and
25.8.degree..+-.0.2.degree. 2.theta., and further wherein the X-ray
powder diffractogram is determined on a diffractometer using
Cu-K.alpha. radiation at a wavelength of 1.5418 .ANG..
2. The crystalline form of claim 1, wherein the crystalline form
further comprises peaks (.degree. 2.theta.) at
6.0.degree..+-.0.2.degree. 2.theta., 6.2.degree..+-.0.2.degree.
2.theta., 8.6.degree..+-.0.2.degree. 2.theta. and
9.6.degree..+-.0.2.degree. 2.theta..
3. A pharmaceutical composition comprising the crystalline form of
claim 1, and at least one pharmaceutically acceptable vehicle.
4. A method for inhibiting spleen tyrosine kinase activity in a
subject in need thereof, comprising administering to the subject a
crystalline Form I of a monomesylate salt of Compound 2:
##STR00150## wherein the crystalline form is characterized by an
X-ray powder diffractogram comprising peaks (.degree.2.theta.) at
17.3.degree..+-.0.2.degree. 2.theta., 17.9.degree..+-.0.2.degree.
2.theta., 19.7.degree..+-.0.2.degree. 2.theta.,
21.6.degree..+-.0.2.degree. 2.theta. and
25.8.degree..+-.0.2.degree. 2.theta., and further wherein the X-ray
powder diffractogram is determined on a diffractometer using
Cu-K.alpha. radiation at a wavelength of 1.5418 .ANG..
5. The method of claim 4, wherein the subject suffers from a
disorder or disease selected from the group consisting of an
inflammatory disorder, an allergic disorder, an autoimmune disease,
and a cancer.
6. The method of claim 4, wherein the subject suffers from is an
inflammatory disorder.
7. The method of claim 4, wherein the subject suffers from a solid
tumor, wherein the solid tumor is from a cancer selected from the
group consisting of pancreatic cancer, urological cancer, bladder
cancer, colorectal cancer, colon cancer, breast cancer, prostate
cancer, renal cancer, hepatocellular cancer, thyroid cancer, gall
bladder cancer, lung cancer, ovarian cancer, cervical cancer,
gastric cancer, endometrial cancer, esophageal cancer, head and
neck cancer, melanoma, neuroendocrine cancer, central nervous
system cancer, a brain turnoff, bone cancer, soft tissue sarcoma,
retinoblastomas, neuroblastomas, peritoneal effusions, malignant
pleural effusions, mesotheliomas, Wilms' tumors, trophoblastic
neoplasms, hemangiopericytomas, Kaposi's sarcomas, myxoid
carcinoma, round cell carcinoma, squamous cell carcinomas,
esophageal squamous cell carcinomas, oral carcinomas, cancers of
the adrenal cortex, and adrenocorticotropic hormone-producing
tumors.
8. The method of claim 7, wherein the lung cancer is non-small cell
lung cancer or small-cell lung cancer.
9. The method of claim 7, wherein the brain tumor is glioma,
anaplastic oligodendroglioma, adult glioblastoma multiforme, or
adult anaplastic astrocytoma.
10. The method of claim 4, wherein the subject suffers from a
disorder or disease selected from the group consisting of small
lymphocytic lymphoma, non-Hodgkin's lymphoma, indolent
non-Hodgkin's lymphoma, refractory indolent non-Hodgkin's lymphoma,
mantle cell lymphoma, follicular lymphoma, lymphoplasmacytic
lymphoma, marginal zone lymphoma, immunoblastic large cell
lymphoma, lymphoblastic lymphoma, splenic marginal zone B-cell
lymphoma, nodal marginal zone lymphoma, extranodal marginal zone
B-cell lymphoma of mucosa-associated lymphoid tissue type,
cutaneous T-cell lymphoma, extranodal T-cell lymphoma, anaplastic
large cell lymphoma, angioimmunoblastic T-cell lymphoma, mycosis
fungoides, B-cell lymphoma, diffuse large B-cell lymphoma,
mediastinal large B-cell lymphoma, intravascular large B-cell
lymphoma, primary effusion lymphoma, small non-cleaved cell
lymphoma, Burkitt's lymphoma, multiple myeloma, plasmacytoma, acute
lymphocytic leukemia, T-cell acute lymphoblastic leukemia, B-cell
acute lymphoblastic leukemia, B-cell prolymphocytic leukemia, acute
myeloid leukemia, chronic lymphocytic leukemia, juvenile
myelomonocytic leukemia, minimal residual disease, hairy cell
leukemia, primary myelofibrosis, secondary myelofibrosis, chronic
myeloid leukemia, myelodysplastic syndrome, myeloproliferative
disease, and Waldestrom's macroglobulinemia.
11. The method of claim 4, wherein the subject suffers from a
disorder or disease selected from the group consisting of systemic
lupus erythematosus, myestenia gravis, Goodpasture's syndrome,
glomerulonephritis, hemorrhage, pulmonary hemorrhage,
atherosclerosis, rheumatoid arthritis, psoriatic arthritis,
monoarticular arthritis, osteoarthritis, gouty arthritis,
spondylitis, Behcet's disease, autoimmune thyroiditis, Reynaud's
syndrome, acute disseminated encephalomyelitis, chronic idiopathic
thrombocytopenic purpura, multiple sclerosis, Sjogren's syndrome,
autoimmune hemolytic anemia, tissue graft rejection, hyperacute
rejection of transplanted organs, allograft rejection,
graft-versus-host disease, diseases involving leukocyte diapedesis,
disease states due to leukocyte dyscrasia and metastasis,
granulocyte transfusion-associated syndromes, cytokine-induced
toxicity, scleroderma, vasculitis, asthma, psoriasis, chronic
inflammatory bowel disease, ulcerative colitis, Crohn's disease,
necrotizing enterocolitis, irritable bowel syndrome,
dermatomyositis, Addison's disease, Parkinson's disease,
Alzheimer's disease, diabetes, type I diabetes mellitus, sepsis,
septic shock, endotoxic shock, gram negative sepsis, gram positive
sepsis, toxic shock syndrome, multiple organ injury syndrome
secondary to septicemia, trauma, hypovolemic shock, allergic
conjunctivitis, vernal conjunctivitis, thyroid-associated
ophthalmopathy, eosinophilic granuloma, eczema, chronic bronchitis,
acute respiratory distress syndrome, allergic rhinitis, coryza, hay
fever, bronchial asthma, silicosis, pulmonary sarcoidosis,
pleurisy, alveolitis, emphysema, pneumonia, bacterial pneumonia,
bronchiectasis, pulmonary oxygen toxicity, reperfusion injury of
the myocardium, brain, or extremities, thermal injury, cystic
fibrosis, keloid formation or scar tissue formation, fever and
myalgias due to infection, brain or spinal cord injury due to minor
trauma, diseases involving leukocyte diapedesis, acute
hypersensitivity, delayed hypersensitivity, urticaria, food
allergies, skin sunburn, inflammatory pelvic disease, urethritis,
uveitis, sinusitis, pneumonitis, encephalitis, meningitis,
myocarditis, nephritis, osteomyelitis, myositis, hepatitis,
alcoholic hepatitis, gastritis, enteritis, contact dermatitis,
atopic dermatitis, gingivitis, appendicitis, pancreatitis,
cholocystitis, polycythemia vera, essential thrombocythemia, and
polycystic kidney disease.
12. The method of claim 4, wherein the subject suffers from a
disorder or disease selected from the group consisting of systemic
lupus erythematosus, myestenia gravis, rheumatoid arthritis, acute
disseminated encephalomyelitis, idiopathic thrombocytopenic
purpura, multiple sclerosis, Sjogren's syndrome, psoriasis,
autoimmune hemolytic anemia, asthma, ulcerative colitis, Crohn's
disease, irritable bowel disease, and chronic obstructive pulmonary
disease.
13. The method of claim 4, wherein the subject suffers from a
disorder or disease selected from the group consisting of asthma,
rheumatoid arthritis, multiple sclerosis, chronic obstructive
pulmonary disease, and systemic lupus erythematosus.
14. The method of claim 4, wherein the subject suffers from a
cancer selected from the group consisting of a hematologic
malignancy and a solid tumor.
15. The method of claim 4, wherein the subject suffers from a
hematologic malignancy selected from the group consisting of
lymphoma, multiple myeloma, and leukemia.
Description
FIELD
The present disclosure relates to compounds and to their use in the
treatment of various diseases, including cancer and inflammatory
conditions. The disclosure also relates to methods for preparation
of the compounds and to pharmaceutical compositions comprising such
compounds.
BACKGROUND
Protein kinases, the largest family of human enzymes, encompass
well over 500 proteins. Spleen Tyrosine Kinase (Syk) is a member of
the Syk family of tyrosine kinases, and is a regulator of early
B-cell development as well as mature B-cell activation, signaling,
and survival.
The inhibition of Syk activity can be useful for the treatment of
allergic disorders, autoimmune diseases and inflammatory diseases
such as: SLE, rheumatoid arthritis, multiple vasculitides,
idiopathic thrombocytopenic purpura (ITP), myasthenia gravis,
allergic rhinitis, chronic obstructive pulmonary disease (COPD),
adult respiratory distress syndrome (ARDs) and asthma. In addition,
Syk has been reported to play an important role in
ligand-independent tonic signaling through the B-cell receptor,
known to be an important survival signal in B-cells. Thus,
inhibition of Syk activity may also be useful in treating certain
types of cancer, including B-cell lymphoma and leukemia. U.S. Pat.
Nos. 8,455,493 and 8,440,667 disclose Syk inhibitors, the
disclosures of which are hereby incorporated by reference in their
entirety.
There is a continued need to provide compounds that are effective
Syk inhibitors, including compounds having desirable
pharmacokinetic properties for use as therapeutics for treating
cancers and other diseases.
SUMMARY
Accordingly, the present disclosure provides compounds that
function as Syk inhibitors. In one embodiment, the disclosure
provides a compound of Formula I:
##STR00002## or a pharmaceutically acceptable salt,
pharmaceutically acceptable co-crystal, pharmaceutically acceptable
ester, stereoisomer, mixture of stereoisomers or tautomer thereof,
wherein:
R.sup.1 is selected from the group consisting of
##STR00003## wherein * indicates the carbon atom of the indicated
phenyl ring of Formula I to which R.sup.1 is attached;
R.sup.2 is H or 2-hydroxyethoxyl;
R.sup.3 is H or methyl; and
R.sup.4 is H or methyl.
Within each of the embodiments described herein comprising a
compound of Formula I, there is a further embodiment wherein each
of R.sup.2, R.sup.3, and R.sup.4 is H. Within each of the
embodiments described herein comprising a compound of Formula I,
there is a another embodiment wherein R.sup.2 is H, R.sup.3 is
methyl, and R.sup.4 is H. Within each of the embodiments described
herein comprising a compound of Formula I, there is also another
embodiment wherein R.sup.2 is H, R.sup.3 is H, and R.sup.4 is
methyl.
Within each of the embodiments described herein comprising a
compound of Formula I, there is still another embodiment wherein
R.sup.2 is 2-hydroxyethoxyl, R.sup.3 is methyl, and R.sup.4 is
H.
Within each of the embodiments described herein comprising a
compound of Formula I, there is still another embodiment wherein
R.sup.2 is 2-hydroxyethoxyl, R.sup.3 is methyl, and R.sup.4 is
H.
Within each of the embodiments described herein comprising a
compound of Formula I, there is still further embodiment wherein
R.sup.2 is 2-hydroxyethoxyl, R.sup.3 is H, and R.sup.4 is
methyl.
Provided herein are also methods of using the compound of Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof, in
the treatment of a disease or condition in a subject, such as a
human. Provided herein are also methods of using the compound of
Formula II, shown below, or a pharmaceutically acceptable salt or
co-crystal thereof, in the treatment of a disease or condition in a
subject, such as a human. Also provided is a compound of Formula I,
or a pharmaceutically acceptable salt or co-crystal thereof, for
use in therapy. Also provided is a compound of Formula II, or a
pharmaceutically acceptable salt or co-crystal thereof, for use in
therapy. Also provided is a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, for use in
the treatment of a disease or condition in a subject, such as a
human. Also provided is a compound of Formula II, or a
pharmaceutically acceptable salt or co-crystal thereof, for use in
the treatment of a disease or condition in a subject, such as a
human. Also provided are uses of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, in the
manufacture of a medicament for the treatment of disease or
condition in a subject, such as a human. Also provided are uses of
the compound of Formula II, or a pharmaceutically acceptable salt
or co-crystal thereof, in the manufacture of a medicament for the
treatment of disease or condition in a subject, such as a human.
Such diseases and conditions include inflammatory disorders,
allergic disorders, autoimmune diseases, or a cancer (including
carcinoma, sarcoma, melanoma, lymphoma and leukemia).
In some instances, the diseases and conditions that may be treated
with the compounds disclosed herein include cancers such as bladder
cancer, breast cancer, colorectal cancer, endometrial cancer,
kidney/renal-cell cancer, lung cancer, pancreatic cancer, prostate
cancer, thyroid cancer, leukemia, melanoma, and non-Hodgkin's
lymphoma.
In some embodiments, the disease is cancer, including a hematologic
malignancy or a solid tumor. In some embodiments, the cancer is
lymphoma, multiple myeloma, or leukemia. In some embodiments, the
hematologic malignancy is leukemia or lymphoma.
In some embodiments, the disclosure provides pharmaceutical
compositions comprising a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, and a
pharmaceutically acceptable vehicle. In other embodiments, the
disclosure provides pharmaceutical compositions comprising a
compound of Formula II, or a pharmaceutically acceptable salt or
co-crystal thereof, and a pharmaceutically acceptable vehicle.
In some embodiments, the disclosure provides pharmaceutical
compositions comprising a therapeutically effective amount of a
compound of Formula I, or a pharmaceutically acceptable salt,
pharmaceutically acceptable co-crystal, pharmaceutically acceptable
ester, stereoisomer, mixture of stereoisomers or tautomer thereof,
and at least one pharmaceutically acceptable vehicle. In other
embodiments, the disclosure provides pharmaceutical compositions
comprising a therapeutically effective amount of a compound of
Formula II, or a pharmaceutically acceptable salt, pharmaceutically
acceptable co-crystal, pharmaceutically acceptable ester,
stereoisomer, mixture of stereoisomers or tautomer thereof, and at
least one pharmaceutically acceptable vehicle. Examples of
pharmaceutically acceptable vehicle may be selected from carriers
and other excipients, adjuvants and the like.
Also provided are methods of treating a disease or condition in a
subject in need thereof by administering to the subject a
therapeutically effective amount of a compound of Formula I, or a
pharmaceutically acceptable salt, pharmaceutically acceptable
co-crystal, pharmaceutically acceptable ester, stereoisomer,
mixture of stereoisomers or tautomer thereof, or a pharmaceutical
composition thereof. In one variation of a method of treating a
disease or condition in a subject in need thereof (e.g., a human in
need thereof), the method comprises administering to the subject a
therapeutically effective amount of a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof. In some
embodiments, the disease or condition is an inflammatory disorder,
an allergic disorder, an autoimmune disease, or a cancer.
Also provided is a method of inhibiting kinase activity of a Syk
kinase polypeptide by contacting the polypeptide with a compound of
Formula I or a pharmaceutically acceptable salt, pharmaceutically
acceptable co-crystal, pharmaceutically acceptable ester,
stereoisomer, mixture of stereoisomers or tautomer thereof. Also
provided is a method of inhibiting kinase activity of a Syk kinase
polypeptide by contacting the polypeptide with a compound of
Formula II or a pharmaceutically acceptable salt, pharmaceutically
acceptable co-crystal, pharmaceutically acceptable ester,
stereoisomer, mixture of stereoisomers or tautomer thereof. In one
aspect is provided a method of inhibiting kinase activity of a Syk
kinase polypeptide by contacting the polypeptide with a compound of
Formula I or a pharmaceutically acceptable salt or co-crystal
thereof. In one aspect, these methods of inhibiting kinase activity
are performed in vitro. In another aspect is provided a method of
inhibiting kinase activity of a Syk kinase polypeptide by
contacting the polypeptide with a compound of Formula II or a
pharmaceutically acceptable salt or co-crystal thereof. In one
aspect, these methods of inhibiting kinase activity are performed
in vitro.
Also provided is a kit that includes a compound of Formula I, or a
pharmaceutically acceptable salt, pharmaceutically acceptable
co-crystal, pharmaceutically acceptable ester, stereoisomer,
mixture of stereoisomers or tautomer thereof. Also provided is a
kit that includes a compound of Formula II, or a pharmaceutically
acceptable salt, pharmaceutically acceptable co-crystal,
pharmaceutically acceptable ester, stereoisomer, mixture of
stereoisomers or tautomer thereof. In one aspect, the kit comprises
a compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof. In a further aspect, the kit comprises a
compound of Formula II, or a pharmaceutically acceptable salt or
co-crystal thereof. The kit may comprise a label and/or
instructions for use of the compound in the treatment of a disease
or condition in a subject (e.g., human) in need thereof. In some
embodiments, the disease or condition may be associated with or
mediated by Syk activity.
Also provided are articles of manufacture that include a compound
of Formula I, or a pharmaceutically acceptable salt,
pharmaceutically acceptable co-crystal, pharmaceutically acceptable
ester, stereoisomer, mixture of stereoisomers or tautomer thereof;
and a container. Also provided are articles of manufacture that
include a compound of Formula II, or a pharmaceutically acceptable
salt, pharmaceutically acceptable co-crystal, pharmaceutically
acceptable ester, stereoisomer, mixture of stereoisomers or
tautomer thereof; and a container. In one aspect, the article of
manufacture comprises a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof. In one
embodiment, the container may be a vial, jar, ampoule, preloaded
syringe, or an intravenous bag. In another aspect, the article of
manufacture comprises a compound of Formula II, or a
pharmaceutically acceptable salt or co-crystal thereof. In one
embodiment, the container may be a vial, jar, ampoule, preloaded
syringe, or an intravenous bag.
In some embodiments, the invention relates to a compound of Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof. In
some embodiments, the invention relates to a compound of Formula
II, or a pharmaceutically acceptable salt or co-crystal
thereof.
Additional aspects and embodiments of this disclosure are described
throughout.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is an XRPD Analysis of Mono MSA Salt Form I of the compound
of Example 2.
FIG. 2 is an NMR Analysis of Mono MSA Salt Form I of the compound
of Example 2.
FIG. 3 is a DSC Analysis of Mono MSA Salt Form I of the compound of
Example 2.
FIG. 4 is a TGA Analysis of Mono MSA Salt Form I of the compound of
Example 2.
FIG. 5 is an XRPD Analysis Mono MSA Salt Form II of the compound of
Example 2.
FIG. 6 is an NMR Analysis Mono MSA Salt Form II of the compound of
Example 2.
FIG. 7 is a DSC Analysis of Mono MSA Salt Form II of the compound
of Example 2.
FIG. 8 is a TGA Analysis of Mono MSA Salt Form II of the compound
of Example 2.
FIG. 9 is an XRPD Analysis of Succinate Form I of the compound of
Example 2.
FIG. 10 is an NMR Analysis of Succinate Form I of the compound of
Example 2.
FIG. 11 is a DSC Analysis of Succinate Form I of the compound of
Example 2.
FIG. 12 is a TGA Analysis of Succinate Form I of the compound of
Example 2.
FIG. 13 is an XRPD Analysis of Succinate Form II of the compound of
Example 2.
FIG. 14 is an NMR Analysis of Succinate Form II of the compound of
Example 2.
FIG. 15 is a DSC Analysis of Succinate Form II of the compound of
Example 2.
FIG. 16 is a TGA Analysis of Succinate Form II of the compound of
Example 2.
DETAILED DESCRIPTION
It has surprisingly been discovered that compounds of Formula I, or
pharmaceutically acceptable salts or co-crystals thereof, possess
advantageous properties, making them attractive compounds for use
as described herein. The compounds, in addition to being Syk
inhibitors, possess desirable solubility and pharmacokinetic
properties. These findings are particularly striking in view of the
properties of comparable parameters of compounds of similar base
structure.
The following description sets forth exemplary methods, parameters
and the like. It should be recognized, however, that such
description is not intended as a limitation on the scope of the
present disclosure but is instead provided as a description of
exemplary embodiments.
Also described for a compound of Formula I are the pharmaceutically
acceptable salts, pharmaceutically acceptable co-crystals,
pharmaceutically acceptable esters, pharmaceutically acceptable
solvates, hydrates, isomers (including optical isomers, racemates,
or other mixtures thereof), tautomers, isotopes, polymorphs, and
pharmaceutically acceptable prodrugs of such compounds.
The compounds of the disclosure may possess an asymmetric center,
and can be produced as a racemic mixture or as individual
enantiomers. The individual enantiomers may be obtained by
asymmetric synthesis or by resolving a racemic or non-racemic
mixture of an intermediate at some appropriate stage of the
synthesis. The individual enantiomers may also be obtained by
resolution of the compound by conventional means, such as
crystallization in the presence of a resolving agent, or
chromatography, using, for example a chiral high pressure liquid
chromatography (HPLC) column. The individual enantiomers as well as
racemic and non-racemic mixtures of enantiomers are within the
scope of the present disclosure, all of which are intended to be
included within the structures depicted in this specification
unless otherwise specifically indicated.
Definitions
As used in the present disclosure, the following words and phrases
are generally intended to have the meanings as set forth below,
except to the extent that the context in which they are used
indicates otherwise.
"Isomers" are different compounds that have the same molecular
formula. Isomers include stereoisomers, enantiomers and
diastereomers.
"Stereoisomers" are isomers that differ only in the way the atoms
are arranged in space.
"Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other. A 1:1 mixture of a
pair of enantiomers is a "racemic" mixture. The term "(.+-.)" is
used to designate a racemic mixture where appropriate.
The absolute stereochemistry is specified according to the Cahn
Ingold Prelog R S system. When the compound is a pure enantiomer
the stereochemistry at each chiral carbon may be specified by
either R or S. Resolved compounds whose absolute configuration is
unknown are designated (+) or (-) depending on the direction
(dextro- or laevorotary) that they rotate the plane of polarized
light at the wavelength of the sodium D line.
The term "therapeutically effective amount" or "pharmaceutically
effective amount" refers to an amount that is sufficient to effect
treatment, as defined below, when administered to a subject (e.g.,
a mammal, such as a human) in need of such treatment. The
therapeutically or pharmaceutically effective amount will vary
depending upon the subject and disease condition being treated, the
weight and age of the subject, the severity of the disease
condition, the manner of administration and the like, which can
readily be determined by one of ordinary skill in the art. For
example, a "therapeutically effective amount" or a
"pharmaceutically effective amount" of a compound of Formula I, or
a pharmaceutically acceptable salt or co-crystal thereof, is an
amount sufficient to modulate Syk expression or activity, and
thereby treat a subject (e.g., a human) suffering an indication, or
to ameliorate or alleviate the existing symptoms of the indication.
For example, a therapeutically or pharmaceutically effective amount
may be an amount sufficient to decrease a symptom of a disease or
condition responsive to inhibition of Syk activity.
The term "polymorph" refers to different crystal structures of a
crystalline compound. The different polymorphs may result from
differences in crystal packing (packing polymorphism) or
differences in packing between different conformers of the same
molecule (conformational polymorphism). It is understood that any
polymorph of a compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, used in the treatment of a
disease or condition as described herein, while possibly providing
varied properties, including pharmacokinetic properties, once
absorbed into the subject, results in the compound of Formula I or
a compound of Formula II, such that the use of a compound of
Formula I or a compound of Formula II encompasses the use of any
polymorph of a compound of Formula I or a compound of Formula II,
respectively, or a pharmaceutically acceptable salt or co-crystal
thereof.
The term "solvate" refers to a complex formed by the combining of a
compound of Formula I or a compound of Formula II and a solvent. It
is understood that any solvate of a compound of Formula I or a
compound of Formula II used in the treatment of a disease or
condition as described herein, while possibly providing varied
properties, including pharmacokinetic properties, once absorbed
into the subject, results in the compound of Formula I or a
compound of Formula II, such that the use of a compound of Formula
I or a compound of Formula II encompasses the use of any solvate of
a compound of Formula I or a compound of Formula II,
respectively.
The term "hydrate" refers to the complex formed by the combining of
a compound of Formula I or a compound of Formula II, or a
pharmaceutically acceptable salt or co-crystal thereof, and water.
It is understood that any hydrate of a compound of Formula I or a
compound of Formula II, or a pharmaceutically acceptable salt or
co-crystal thereof, used in the treatment of a disease or condition
as described herein, while possibly providing varied properties,
including pharmacokinetic properties, once absorbed into the
subject, results in the compound of Formula I or of Formula II,
such that the use of a compound of Formula I or Formula II
encompasses the use of any hydrate of a compound of Formula I or
Formula II, respectively.
The term "prodrug" refers to a compound derived from or readily
converted to a compound of Formula I or of Formula II that include
chemical groups which, in vivo, can be converted and/or can be
split off from the remainder of the molecule to provide a compound
of Formula I or of Formula II or active moiety of the drug, or a
pharmaceutically acceptable salt or co-crystal thereof or a
biologically active metabolite thereof. It is understood that any
prodrug of a compound of Formula I or of Formula II used in the
treatment of a disease or condition as described herein, while
possibly providing varied properties, including pharmacokinetic
properties, once absorbed into the subject, results in the compound
of Formula I or Formula II, such that the use of a compound of
Formula I or Formula II encompasses the use of any prodrug of a
compound of Formula I or Formula II, respectively. Prodrugs can,
for example, be produced by replacing functionalities present in
the compounds of the invention with appropriate moieties which are
metabolized in vivo to form a compound of the invention. The design
of prodrugs is well-known in the art, as discussed in Bundgaard,
Design of Prodrugs 1985 (Elsevier), The Practice of Medicinal
Chemistry 2003, 2nd Ed, 561-585 and Leinweber, Drug Metab. Res.
1987, 18: 379.
Examples of prodrugs of compounds of the invention are esters and
amides of the compounds of the invention. For example, where the
compound of the invention contains an alcohol group (--OH), the
hydrogen atom of the alcohol group may be replaced in order to form
an ester (e.g. the hydrogen atom may be replaced by
--C(O)C.sub.1-6alkyl. Where the compound of the invention contains
a primary or secondary amino group, one or more hydrogen atoms of
the amino group may be replaced in order to form an amide (e.g. one
or more hydrogen atoms may be replaced by C(O)C.sub.1-6alkyl).
Also provided herein are isotopically labeled forms of compounds
detailed herein. Isotopically labeled compounds have structures
depicted by the formulas given herein except that one or more atoms
are replaced by an atom having a selected atomic mass or mass
number. Examples of isotopes that can be incorporated into
compounds of the disclosure include isotopes of hydrogen, carbon,
nitrogen, oxygen, phosphorous, fluorine and chlorine, such as, but
not limited to .sup.2H (deuterium, D), .sup.3H (tritium), .sup.11C,
.sup.13C, .sup.14C, .sup.15N, .sup.18F, .sup.31P, .sup.32P,
.sup.35S, .sup.36Cl and .sup.125I. Various isotopically labeled
compounds of the present disclosure, for example those into which
radioactive isotopes such as .sup.3H, .sup.13C and .sup.14C are
incorporated, are provided. Such isotopically labeled compounds may
be useful in metabolic studies, reaction kinetic studies, detection
or imaging techniques, such as positron emission tomography (PET)
or single-photon emission computed tomography (SPECT) including
drug or substrate tissue distribution assays or in radioactive
treatment of subjects (e.g. humans). Also provided for isotopically
labeled compounds described herein are any pharmaceutically
acceptable salts, pharmaceutically acceptable esters,
pharmaceutically acceptable solvates, hydrates, enantiomers,
mixture of enantiomers, tautomers, polymorphs, and pharmaceutically
acceptable prodrugs thereof.
The disclosure also includes the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, in which
from 1 to n hydrogens attached to a carbon atom is/are replaced by
deuterium, in which n is the number of hydrogens in the molecule.
Such compounds may exhibit increased resistance to metabolism and
are thus useful for increasing the half life of the compound of
Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, when administered to a mammal. See, for example, Foster,
"Deuterium Isotope Effects in Studies of Drug Metabolism", Trends
Pharmacol. Sci. 5(12):524-527 (1984). Such compounds are
synthesized by means well known in the art, for example by
employing starting materials in which one or more hydrogens have
been replaced by deuterium.
Deuterium labeled or substituted therapeutic compounds of the
disclosure may have improved DMPK (drug metabolism and
pharmacokinetics) properties, relating to distribution, metabolism
and excretion (ADME). Substitution with heavier isotopes such as
deuterium may afford certain therapeutic advantages resulting from
greater metabolic stability, for example increased in vivo
half-life, reduced dosage requirements and/or an improvement in
therapeutic index. An .sup.18F labeled compound may be useful for
PET or SPECT studies. Isotopically labeled compounds of this
disclosure and prodrugs thereof can generally be prepared by
carrying out the procedures disclosed in the schemes or in the
examples and preparations described below by substituting a readily
available isotopically labeled reagent for a non-isotopically
labeled reagent. It is understood that deuterium in this context is
regarded as a substituent in the compound of Formula I.
The concentration of such a heavier isotope, specifically
deuterium, may be defined by an isotopic enrichment factor. In the
compounds of this disclosure any atom not specifically designated
as a particular isotope is meant to represent any stable isotope of
that atom. Unless otherwise stated, when a position is designated
specifically as "H" or "hydrogen", the position is understood to
have hydrogen at its natural abundance isotopic composition.
Accordingly, in the compounds of this disclosure any atom
specifically designated as a deuterium (D) is meant to represent
deuterium.
The term "inhibition" indicates a decrease, such as a significant
decrease, in the baseline activity of a biological activity or
process. "Inhibition of Syk activity" refers to a decrease in Syk
activity as a direct or indirect response to the presence of a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, relative to the activity of Syk in the absence
of such compound or a pharmaceutically acceptable salt or
co-crystal thereof. The decrease in activity may be due to the
direct interaction of the compound with Syk, or due to the
interaction of the compound(s) described herein with one or more
other factors that in turn affect Syk activity. For example, the
presence of the compound(s) may decrease Syk activity by directly
binding to the Syk, by causing (directly or indirectly) another
factor to decrease Syk activity, or by (directly or indirectly)
decreasing the amount of Syk present in the cell or organism. In
some embodiments, the inhibition of Syk activity may be compared in
the same subject prior to treatment, or other subjects not
receiving the treatment.
Inhibition of Syk activity also refers to observable inhibition of
Syk activity in a standard biochemical assay for Syk activity, such
as the ATP hydrolysis assay described in Example 12 below.
In some embodiments, the compound described herein, e.g. a compound
of Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, inhibits Syk kinase activity with an IC.sub.50 value less
than or equal to 1 micromolar, such as 0.1 nM to 1 .mu.M or 1 nM to
1 .mu.M. In some embodiments, the compound or a pharmaceutically
acceptable salt or co-crystal thereof has an IC.sub.50 value less
than or equal to less than 500 nanomolar, such as 0.1 nM to 500 nM
or 1 nM to 500 nM. In some embodiments, the compound or a
pharmaceutically acceptable salt or co-crystal thereof has an
IC.sub.50 value less than or equal to less than 200 nanomolar, such
as 0.1 nM to 200 nM or 1 nM to 200 nM. In some embodiments, the
compound or a pharmaceutically acceptable salt or co-crystal
thereof has an IC.sub.50 value less than or equal to less than 100
nanomolar, such as 0.1 nM to 100 nM or 1 nM to 100 nM. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has an IC.sub.50 value less than or equal to 50
nanomolar, such as 0.1 nM to 50 nM or 1 nM to 50 nM. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has an IC.sub.50 value less than or equal to 20
nanomolar, such as 0.1 nM to 20 nM or 1 nM to 20 nM. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has an IC.sub.50 value less than or equal to 10
nanomolar, such as 0.1 nM to 10 nM or 1 nM to 10 nM. In some
embodiments, the IC.sub.50 value is measured as described in the
assay of Example 12.
"Inhibition of B-cell activity" refers to a decrease in B-cell
activity as a direct or indirect response to the presence of a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, relative to the activity of B-cells in the
absence of such compound or a pharmaceutically acceptable salt or
co-crystal thereof. The decrease in activity may be due to the
direct interaction of the compound with Syk or with one or more
other factors that in turn affect B-cell activity.
Inhibition of B-cell activity also refers to observable inhibition
of CD86 expression in a standard assay. In some embodiments, the
compound described herein has an IC.sub.50 value less than or equal
to 10 micromolar, such as 1 nM to 10 .mu.M or 10 nM to 10 .mu.M. In
some embodiments, the compound has an IC.sub.50 value less than or
equal to less than 1 micromolar, such as 1 nM to 1 .mu.M or 10 nM
to 1 .mu.M. In some embodiments, the compound has an IC.sub.50
value less than or equal to 500 nanomolar, such as 1 nM to 500 nM
or 10 nM to 500 nM.
"B cell activity" also includes activation, redistribution,
reorganization, or capping of one or more various B cell membrane
receptors, or membrane-bound immunoglobulins, e.g., IgM, IgG, and
IgD. Most B cells also have membrane receptors for the Fc portion
of IgG in the form of either antigen-antibody complexes or
aggregated IgG. B cells also carry membrane receptors for the
activated components of complement, e.g., C3b, C3d, C4, and Clq.
These various membrane receptors and membrane-bound immunoglobulins
have membrane mobility and can undergo redistribution and capping
that can initiate signal transduction.
B cell activity also includes the synthesis or production of
antibodies or immunoglobulins. Immunoglobulins are synthesized by
the B cell series and have common structural features and
structural units. Five immunoglobulin classes, i.e., IgG, IgA, IgM,
IgD, and IgE, are recognized on the basis of structural differences
of their heavy chains including the amino acid sequence and length
of the polypeptide chain. Antibodies to a given antigen may be
detected in all or several classes of immunoglobulins or may be
restricted to a single class or subclass of immunoglobulin.
Autoantibodies or autoimmune antibodies may likewise belong to one
or several classes of immunoglobulins. For example, rheumatoid
factors (antibodies to IgG) are most often recognized as an IgM
immunoglobulin, but can also consist of IgG or IgA.
In addition, B cell activity also is intended to include a series
of events leading to B cell clonal expansion (proliferation) from
precursor B lymphocytes and differentiation into
antibody-synthesizing plasma cells which takes place in conjunction
with antigen-binding and with cytokine signals from other
cells.
"Inhibition of B-cell proliferation" refers to inhibition of
proliferation of abnormal B-cells, such as cancerous B-cells, e.g.
lymphoma B-cells and/or inhibition of normal, non-diseased B-cells.
The term "inhibition of B-cell proliferation" indicates any
significant decrease in the number of B-cells, either in vitro or
in vivo. Thus an inhibition of B-cell proliferation in vitro would
be any significant decrease in the number of B-cells in an in vitro
sample contacted with a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof as compared
to a matched sample not contacted with the compound(s).
Inhibition of B-cell proliferation also refers to observable
inhibition of B-cell proliferation in a standard thymidine
incorporation assay for B-cell proliferation, e.g. such assay as
known in the art. In some embodiments, the compounds described
herein, e.g. a compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, has an IC.sub.50 value less
than or equal to 10 micromolar, such as 1 nM to 10 .mu.M or 10 nM
to 10 .mu.M. In some embodiments, the compound or a
pharmaceutically acceptable salt or co-crystal thereof has an
IC.sub.50 value less than or equal to less than 1 micromolar, such
as 1 nM to 1 .mu.M or 10 nM to 1 .mu.M. In some embodiments, the
compound or a pharmaceutically acceptable salt or co-crystal
thereof has an IC.sub.50 value less than or equal to 500 nanomolar,
such as 1 nM to 500 nM or 10 nM to 500 nM. In some embodiments, the
compound or a pharmaceutically acceptable salt or co-crystal
thereof has an IC.sub.50 value less than or equal to 200 nanomolar,
such as 1 nM to 200 nM or 10 nM to 200 nM. In some embodiments, the
compound or a pharmaceutically acceptable salt or co-crystal
thereof has an IC.sub.50 value less than or equal to 100 nanomolar,
such as 1 nM to 100 nM or 10 nM to 100 nM.
The "reduction in basophil activation" refers to the ability of
compounds as described herein to reduce the activation of
basophils. Basophil activation is involved, for example, in
inflammatory and autoimmune diseases as described herein, and the
reduction of activation of basophils is desired in compounds as
described herein, e.g. a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof. The
activation of basophils can be assessed by the measurement of CD63
expression by basophils, such as by a CD63 human whole blood
basophil cellular assay (25% blood), e.g. such as the assay
described in Example 9 below.
In some embodiments, the compound described herein e.g. a compound
of Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, has an EC.sub.50 value in a suitable CD63 assay of less
than or equal to 10 micromolar, such as 1 nM to 10 .mu.M or 10 nM
to 10 .mu.M. In some embodiments, the compound or a
pharmaceutically acceptable salt or co-crystal thereof, has an
EC.sub.50 value less than or equal to less than 1 micromolar, such
as 1 nM to 1 .mu.M or 10 nM to 1 .mu.M. In some embodiments, the
compound or a pharmaceutically acceptable salt or co-crystal
thereof has an EC.sub.50 value less than or equal to 500 nanomolar,
such as 1 nM to 500 nM or 10 nM to 500 nM. In some embodiments, the
compound or a pharmaceutically acceptable salt or co-crystal
thereof has an EC.sub.50 value less than or equal to 200 nanomolar,
such as 1 nM to 200 nM or 10 nM to 200 nM. In some embodiments, the
compound or a pharmaceutically acceptable salt or co-crystal
thereof has an EC.sub.50 value less than or equal to 150 nanomolar,
such as 1 nM to 150 nM or 10 nM to 150 nM. In some embodiments, the
compound or a pharmaceutically acceptable salt or co-crystal
thereof has an IC.sub.50 value less than or equal to 100 nanomolar,
such as 1 nM to 100 nM or 10 nM to 100 nM. In some embodiments, the
compound or a pharmaceutically acceptable salt or co-crystal
thereof has an EC.sub.50 value less than or equal to 75 nanomolar,
such as 1 nM to 75 nM or 10 nM to 75 nM. In some embodiments, the
EC.sub.50 value is measured as described in the assay of Example
9.
The "kinetic solubility" refers to an assessment of the solubility
of a compound in a suitable buffer, such as phosphate buffer at pH
7.4, at a given temperature, for example at 37.degree. C. In one
instance, kinetic solubility is measured at 37.degree. C. in
phosphate buffer at pH 7.4, such as by the assay as described in
Example 10.
In some embodiments, the compounds described herein, e.g. a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, has a kinetic solubility at 37.degree. C. in
phosphate buffer at pH 7.4 of greater than or equal to 10 .mu.M,
such as 10 .mu.M to 500 .mu.M or 10 .mu.M to 250 .mu.M. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has a kinetic solubility at 37.degree. C. in
phosphate buffer at pH 7.4 of greater than or equal to 20 .mu.M,
such as 20 .mu.M to 500 .mu.M or 20 .mu.M to 250 .mu.M. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has a kinetic solubility at 37.degree. C. in
phosphate buffer at pH 7.4 of greater than or equal to 30 .mu.M,
such as 30 .mu.M to 500 .mu.M or 30 .mu.M to 250 .mu.M. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has a kinetic solubility at 37.degree. C. in
phosphate buffer at pH 7.4 of greater than or equal to 40 .mu.M,
such as 40 .mu.M to 500 .mu.M or 40 .mu.M to 250 .mu.M. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has a kinetic solubility at 37.degree. C. in
phosphate buffer pH 7.4 of greater than or equal to 50 .mu.M, such
as 50 .mu.M to 500 .mu.M or 50 .mu.M to 250 .mu.M. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has a kinetic solubility at 37.degree. C. in
phosphate buffer at pH 7.4 of greater than or equal to 60 .mu.M,
such as 60 .mu.M to 500 .mu.M or 60 .mu.M to 250 .mu.M. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has a kinetic solubility at 37.degree. C. in
phosphate buffer at pH 7.4 of greater than or equal to 70 .mu.M,
such as 70 .mu.M to 500 .mu.M or 70 .mu.M to 250 .mu.M. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has a kinetic solubility at 37.degree. C. in
phosphate buffer at pH 7.4 of greater than or equal to 80 .mu.M,
such as 80 .mu.M to 500 .mu.M or 80 .mu.M to 250 .mu.M. In some
embodiments, the compound or a pharmaceutically acceptable salt or
co-crystal thereof has a kinetic solubility at 37.degree. C. in
phosphate buffer at pH 7.4 of greater than or equal to 90 .mu.M,
such as 90 .mu.M to 500 .mu.M or 90 .mu.M to 250 .mu.M. In some
embodiments, the kinetic solubility is measured by the assay as
described in Example 10.
The "human hepatocyte stability" is a measure of the stability of
the compounds to metabolism by human hepatocytes, and is assessed
as the predicted hepatic plasma clearance of the compounds in
L/hr/kg. The predicted hepatocyte clearance can be measured, for
example, by the assay described in Example 11.
In some embodiments, the compounds described herein, e.g. a
compound of Formula I or of Formula II has a predicted hepatic
plasma clearance of less than or equal to 0.50 L/hr/kg, such as
0.005 L/hr/kg to 0.50 L/hr/kg or 0.01 L/hr/kg to 0.50 L/hr/kg. In
some embodiments, the compound has a predicted hepatic plasma
clearance of less than or equal to 0.40 L/hr/kg, such as 0.005
L/hr/kg to 0.40 L/hr/kg or 0.01 L/hr/kg to 0.40 L/hr/kg. In some
embodiments, the compound has a predicted hepatic plasma clearance
of less than or equal to 0.30 L/hr/kg, such as 0.005 L/hr/kg to
0.30 L/hr/kg or 0.01 L/hr/kg to 0.30 L/hr/kg. In some embodiments,
the compound has a predicted hepatic plasma clearance of less than
or equal to 0.20 L/hr/kg, such as 0.005 L/hr/kg to 0.20 L/hr/kg or
0.01 L/hr/kg to 0.20 L/hr/kg. In some embodiments, the compound has
a predicted hepatic plasma clearance of less than or equal to 0.10
L/hr/kg, such as 0.005 L/hr/kg to 0.10 L/hr/kg or 0.01 L/hr/kg to
0.10 L/hr/kg. In some embodiments, the compound has a predicted
hepatic plasma clearance of less than or equal to 0.09 L/hr/kg,
such as 0.005 L/hr/kg to 0.09 L/hr/kg or 0.01 L/hr/kg to 0.09
L/hr/kg. In some embodiments, the compound has a predicted hepatic
plasma clearance of less than or equal to 0.08 L/hr/kg, such as
0.005 L/hr/kg to 0.08 L/hr/kg or 0.01 L/hr/kg to 0.08 L/hr/kg. In
some embodiments, the compound has a predicted hepatic plasma
clearance of less than or equal to 0.07 L/hr/kg, such as 0.005
L/hr/kg to 0.07 L/hr/kg or 0.01 L/hr/kg to 0.07 L/hr/kg. In some
embodiments, the compound has a predicted hepatic plasma clearance
of or less than or equal to 0.06 L/hr/kg, such as 0.005 L/hr/kg to
0.06 L/hr/kg or 0.01 L/hr/kg to 0.06 L/hr/kg. In some embodiments,
the predicted hepatocyte clearance is measured by the assay
described in Example 11.
An "allergy" or "allergic disorder" refers to acquired
hypersensitivity to a substance (allergen). Allergic conditions
include eczema, allergic rhinitis or coryza, hay fever, bronchial
asthma, urticaria (hives) and food allergies, and other atopic
conditions.
"Asthma" refers to a disorder of the respiratory system
characterized by inflammation, narrowing of the airways and
increased reactivity of the airways to inhaled agents. Asthma is
frequently, although not exclusively associated with atopic or
allergic symptoms.
By "significant" is meant any detectable change that is
statistically significant in a standard parametric test of
statistical significance such as Student's T-test, where
p<0.05.
A "disease responsive to inhibition of Syk activity" is a disease
in which inhibiting Syk kinase provides a therapeutic benefit such
as an amelioration of symptoms, decrease in disease progression,
delay of disease onset, or inhibition of aberrant activity of
certain cell-types (monocytes, B-cells, and mast cells).
"Subject" refers to an animal, such as a mammal, that has been or
will be the object of treatment, observation or experiment. The
methods described herein may be useful in both human therapy and
veterinary applications. In some embodiments, the subject is a
mammal; in some embodiments the subject is human; and in some
embodiments the subject is chosen from cats and dogs. "Subject in
need thereof" or "human in need thereof" refers to a subject, such
as a human, who may have or is suspected to have diseases or
conditions that would benefit from certain treatment; for example
treatment with a compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, as described herein. This
includes a subject who may be determined to be at risk of or
susceptible to such diseases or conditions, such that treatment
would prevent the disease or condition from developing.
"Treatment" or "treating" is an approach for obtaining beneficial
or desired results including clinical results. Beneficial or
desired clinical results may include one or more of the
following:
(i) inhibiting the disease or condition (e.g., decreasing one or
more symptoms resulting from the disease or condition, and/or
diminishing the extent of the disease or condition);
(ii) slowing or arresting the development of one or more clinical
symptoms associated with the disease or condition (e.g.,
stabilizing the disease or condition, preventing or delaying the
worsening or progression of the disease or condition, and/or
preventing or delaying the spread (e.g., metastasis) of the disease
or condition); and/or
(iii) relieving the disease, that is, causing the regression of
clinical symptoms (e.g., ameliorating the disease state, providing
partial or total remission of the disease or condition, enhancing
effect of another medication, delaying the progression of the
disease, increasing the quality of life, and/or prolonging
survival).
"Delaying" the development of a disease or condition means to
defer, hinder, slow, retard, stabilize, and/or postpone development
of the disease or condition. This delay can be of varying lengths
of time, depending on the history of the disease or condition,
and/or subject being treated. A method that "delays" development of
a disease or condition is a method that reduces probability of
disease or condition development in a given time frame and/or
reduces the extent of the disease or condition in a given time
frame, when compared to not using the method. Such comparisons are
typically based on clinical studies, using a statistically
significant number of subjects. Disease or condition development
can be detectable using standard methods, such as routine physical
exams, mammography, imaging, or biopsy. Development may also refer
to disease or condition progression that may be initially
undetectable and includes occurrence, recurrence, and onset.
In many cases, the compounds of this disclosure are capable of
forming acid and/or base salts by virtue of the presence of amino
and/or carboxyl groups or groups similar thereto.
"Pharmaceutically acceptable salts" include, for example, salts
with inorganic acids and salts with an organic acid. Examples of
salts may include hydrochloride, phosphate, diphosphate,
hydrobromide, sulfate, sulfinate, nitrate, malate, maleate,
fumarate, tartrate, succinate, citrate, acetate, lactate,
methanesulfonate (mesylate), benzenesuflonate (besylate),
p-toluenesulfonate (tosylate), 2-hydroxyethylsulfonate, benzoate,
salicylate, stearate, and alkanoate (such as acetate,
HOOC--(CH.sub.2).sub.n--COOH where n is 0-4). In addition, if the
compounds described herein are obtained as an acid addition salt,
the free base can be obtained by basifying a solution of the acid
salt. Conversely, if the product is a free base, an addition salt,
particularly a pharmaceutically acceptable addition salt, may be
produced by dissolving the free base in a suitable organic solvent
and treating the solution with an acid, in accordance with
conventional procedures for preparing acid addition salts from base
compounds. Those skilled in the art will recognize various
synthetic methodologies that may be used to prepare nontoxic
pharmaceutically acceptable addition salts.
The compound of Formula I or the compound of Formula II can also be
a pharmaceutically acceptable co-crystal or a co-crystal salt. The
"co-crystal" or "co-crystal salt" as used herein means a
crystalline material composed of two or more unique solids at room
temperature, each of which has distinctive physical characteristics
such as structure, melting point, and heats of fusion,
hygroscopicity, solubility, and stability. A co-crystal or a
co-crystal salt can be produced according to a per se known
co-crystallization method. The terms co-crystal (or cocrystal) or
co-crystal salt also refer to a multicomponent system in which
there exists a host API (active pharmaceutical ingredient) molecule
or molecules, such as a compound of Formula I, and a guest (or
co-former) molecule or molecules. In particular embodiments said
pharmaceutically acceptable co-crystal of the compound of Formula I
or of the compound of Formula II with a co-former molecule is in a
crystalline form selected from a malonic acid co-crystal, a
succinic acid co-crystal, a decanoic acid co-crystal, a salicylic
acid co-crystal, a vanillic acid co-crystal, a maltol co-crystal,
or a glycolic acid co-crystal. Co-crystals may have improved
properties as compared to the parent form (i.e., the free molecule,
zwitter ion, etc.) or a salt of the parent compound. Improved
properties can include increased solubility, increased dissolution,
increased bioavailability, increased dose response, decreased
hygroscopicity, a crystalline form of a normally amorphous
compound, a crystalline form of a difficult to salt or unsaltable
compound, decreased form diversity, more desired morphology, and
the like.
The term "crystal forms" and related terms herein refer to the
various crystalline modifications of a given substance, including,
but not limited to, polymorphs, solvates, hydrates, co-crystals,
and other molecular complexes, as well as salts, solvates of salts,
hydrates of salts, other molecular complexes of salts, and
polymorphs thereof. Crystal forms of a substance can be obtained by
a number of methods, as known in the art. Such methods include, but
are not limited to, melt recrystallization, melt cooling, solvent
recrystallization, recrystallization in confined spaces such as,
e.g., in nanopores or capillaries, recrystallization on surfaces or
templates, such as, e.g., on polymers, recrystallization in the
presence of additives, such as, e.g., co-crystal counter-molecules,
desolvation, dehydration, rapid evaporation, rapid cooling, slow
cooling, vapor diffusion, sublimation, grinding and solvent-drop
grinding.
As used herein, "pharmaceutically acceptable excipient" is a
pharmaceutically acceptable vehicle that includes, without
limitation, any and all carriers, solvents, dispersion media,
coatings, antibacterial and antifungal agents, isotonic and
absorption delaying agents and the like. The use of such media and
agents for pharmaceutically active substances is well known in the
art. Except insofar as any conventional media or agent is
incompatible with the active ingredient, its use in the therapeutic
compositions is contemplated. Supplementary active ingredients can
also be incorporated into the compositions.
The term "carrier" refers to an excipient or vehicle that includes
without limitation diluents, disintegrants, precipitation
inhibitors, surfactants, glidants, binders, lubricants, and the
like with which the compound is administered. Carriers are
generally described herein and also in "Remington's Pharmaceutical
Sciences" by E. W. Martin. Examples of carriers include, but are
not limited to, aluminum monostearate, aluminum stearate,
carboxymethylcellulose, carboxymethylcellulose sodium,
crospovidone, glyceryl isostearate, glyceryl monostearate,
hydroxyethyl cellulose, hydroxyethyl cellulose, hydroxymethyl
cellulose, hydroxyoctacosanyl hydroxystearate, hydroxypropyl
cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,
lactose, lactose monohydrate, magnesium stearate, mannitol,
microcrystalline cellulose, poloxamer 124, poloxamer 181, poloxamer
182, poloxamer 188, poloxamer 237, poloxamer 407, povidone, silicon
dioxide, colloidal silicon dioxide, silicone, silicone adhesive
4102, and silicone emulsion. It should be understood, however, that
the carriers selected for the pharmaceutical compositions, and the
amounts of such carriers in the composition, may vary depending on
the method of formulation (e.g., dry granulation formulation, solid
dispersion formulation).
The term "diluent" generally refers to a substance that is used to
dilute the compound of interest prior to delivery. Diluents can
also serve to stabilize compounds. Examples of diluents may include
starch, saccharides, disaccharides, sucrose, lactose,
polysaccharides, cellulose, cellulose ethers, hydroxypropyl
cellulose, sugar alcohols, xylitol, sorbitol, maltitol,
microcrystalline cellulose, calcium or sodium carbonate, lactose,
lactose monohydrate, dicalcium phosphate, cellulose, compressible
sugars, dibasic calcium phosphate dehydrate, mannitol,
microcrystalline cellulose, and tribasic calcium phosphate.
The term "disintegrant" generally refers to a substance which, upon
addition to a solid preparation, facilitates its break-up or
disintegration after administration and permits the release of an
active ingredient as efficiently as possible to allow for its rapid
dissolution. Examples of disintegrants may include maize starch,
sodium starch glycolate, croscarmellose sodium, crospovidone,
microcrystalline cellulose, modified corn starch, sodium
carboxymethyl starch, povidone, pregelatinized starch, and alginic
acid.
The term "precipitation inhibitors" generally refers to a substance
that prevents or inhibits precipitation of the active agent from a
supersaturated solution. One example of a precipitation inhibitor
includes hydroxypropylmethylcellulose (HPMC).
The term "surfactants" generally refers to a substance that lowers
the surface tension between a liquid and a solid that could improve
the wetting of the active agent or improve the solubility of the
active agent. Examples of surfactants include poloxamer and sodium
lauryl sulfate.
The term "glidant" generally refers to substances used in tablet
and capsule formulations to improve flow-properties during tablet
compression and to produce an anti-caking effect. Examples of
glidants may include colloidal silicon dioxide, talc, fumed silica,
starch, starch derivatives, and bentonite.
The term "binder" generally refers to any pharmaceutically
acceptable film which can be used to bind together the active and
inert components of the carrier together to maintain cohesive and
discrete portions. Examples of binders may include
hydroxypropylcellulose, hydroxypropylmethylcellulose, povidone,
copovidone, and ethyl cellulose.
The term "lubricant" generally refers to a substance that is added
to a powder blend to prevent the compacted powder mass from
sticking to the equipment during the tableting or encapsulation
process. A lubricant can aid the ejection of the tablet form the
dies, and can improve powder flow. Examples of lubricants may
include magnesium stearate, stearic acid, silica, fats, calcium
stearate, polyethylene glycol, sodium stearyl fumarate, or talc;
and solubilizers such as fatty acids including lauric acid, oleic
acid, and C.sub.8/C.sub.10 fatty acid.
Compounds
Compounds are provided here and elsewhere throughout, such as in
the Summary and in the Examples.
The compounds provided herein are named using ChemBioDraw Ultra
12.0, and one skilled in the art understands that the compound
structure may be named or identified using other commonly
recognized nomenclature systems and symbols including CAS and
IUPAC.
In some embodiments of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, R.sup.2 is
H. In some embodiments of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, R.sup.2 is
2-hydroxyethoxyl.
In some embodiments of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, R.sup.3 is
H. In some embodiments of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, R.sup.3 is
methyl.
In some embodiments of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, R.sup.4 is
H or methyl. In some embodiments of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, R.sup.4 is
H. In some embodiments of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, R.sup.4 is
methyl.
In some embodiments of the compound of Formula I, or a
pharmaceutically acceptable salt thereof, R.sup.1 is selected from
the group consisting of
##STR00004## In some embodiments, R.sup.1 is
##STR00005## In some embodiments, R.sup.1 is
##STR00006## In some embodiments, R.sup.1 is
##STR00007## In some embodiments, R.sup.1 is
##STR00008##
Separate embodiments herein, each providing a compound of Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof, in
which R.sup.1 is selected from the group consisting of
##STR00009## include embodiments A-1 through A-27 wherein R.sup.2,
R.sup.3, and R.sup.4 are as defined in Table A for each of the
embodiments.
TABLE-US-00001 TABLE A Embodiment No. R.sup.1 selected from R.sup.2
R.sup.3 R.sup.4 A-1 ##STR00010## H or 2- hydroxyethoxyl H or methyl
H or methyl A-2 ##STR00011## H or 2- hydroxyethoxyl H H or methyl
A-3 ##STR00012## H or 2- hydroxyethoxyl methyl H or methyl A-4
##STR00013## H or 2- hydroxyethoxyl H or methyl H A-5 ##STR00014##
H or 2- hydroxyethoxyl H or methyl Methyl A-6 ##STR00015## H or 2-
hydroxyethoxyl H H A-7 ##STR00016## H or 2- hydroxyethoxyl H methyl
A-8 ##STR00017## H or 2- hydroxyethoxyl methyl H A-9 ##STR00018## H
or 2- hydroxyethoxyl methyl methyl A-10 ##STR00019## H H or methyl
H or methyl A-11 ##STR00020## H H H or methyl A-12 ##STR00021## H
methyl H or methyl A-13 ##STR00022## H H or methyl H A-14
##STR00023## H H or methyl methyl A-15 ##STR00024## H H H A-16
##STR00025## H methyl H A-17 ##STR00026## H H methyl A-18
##STR00027## H methyl methyl A-19 ##STR00028## 2-hydroxyethoxyl H
or methyl H or methyl A-20 ##STR00029## 2-hydroxyethoxyl H H or
methyl A-21 ##STR00030## 2-hydroxyethoxyl methyl H or methyl A-22
##STR00031## 2-hydroxyethoxyl H or methyl H A-23 ##STR00032##
2-hydroxyethoxyl H or methyl methyl A-24 ##STR00033##
2-hydroxyethoxyl H H A-25 ##STR00034## 2-hydroxyethoxyl methyl H
A-26 ##STR00035## 2-hydroxyethoxyl H methyl A-27 ##STR00036##
2-hydroxyethoxyl methyl methyl
Separate embodiments herein, each providing a compound of Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof, in
which R.sup.1 is
##STR00037## include embodiments B-1 through B-27 wherein R.sup.2,
R.sup.3, and R.sup.4 are as defined in Table B for each of the
embodiments.
TABLE-US-00002 TABLE B Embodiment No. R.sup.1 R.sup.2 R.sup.3
R.sup.4 B-1 ##STR00038## H or 2- hydroxyethoxyl H or methyl H or
methyl B-2 ##STR00039## H or 2- hydroxyethoxyl H H or methyl B-3
##STR00040## H or 2- hydroxyethoxyl methyl H or methyl B-4
##STR00041## H or 2- hydroxyethoxyl H or methyl H B-5 ##STR00042##
H or 2- hydroxyethoxyl H or methyl Methyl B-6 ##STR00043## H or 2-
hydroxyethoxyl H H B-7 ##STR00044## H or 2- hydroxyethoxyl H methyl
B-8 ##STR00045## H or 2- hydroxyethoxyl methyl H B-9 ##STR00046## H
or 2- hydroxyethoxyl methyl methyl B-10 ##STR00047## H H or methyl
H or methyl B-11 ##STR00048## H H H or methyl B-12 ##STR00049## H
methyl H or methyl B-13 ##STR00050## H H or methyl H B-14
##STR00051## H H or methyl methyl B-15 ##STR00052## H H H B-16
##STR00053## H methyl H B-17 ##STR00054## H H methyl B-18
##STR00055## H methyl methyl B-19 ##STR00056## 2-hydroxyethoxyl H
or methyl H or methyl B-20 ##STR00057## 2-hydroxyethoxyl H H or
methyl B-21 ##STR00058## 2-hydroxyethoxyl methyl H or methyl B-22
##STR00059## 2-hydroxyethoxyl H or methyl H B-23 ##STR00060##
2-hydroxyethoxyl H or methyl methyl B-24 ##STR00061##
2-hydroxyethoxyl H H B-25 ##STR00062## 2-hydroxyethoxyl methyl H
B-26 ##STR00063## 2-hydroxyethoxyl H methyl B-27 ##STR00064##
2-hydroxyethoxyl methyl methyl
Separate embodiments herein, each providing a compound of Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof, in
which R.sup.1 is,
##STR00065## include embodiments C-1 through C-27 wherein R.sup.2,
R.sup.3, and R.sup.4 are as defined in Table C for each of the
embodiments.
TABLE-US-00003 TABLE C Embodiment No. R.sup.1 R.sup.2 R.sup.3
R.sup.4 C-1 ##STR00066## H or 2- hydroxy- ethoxyl H or methyl H or
methyl C-2 ##STR00067## H or 2- hydroxy- ethoxyl H H or methyl C-3
##STR00068## H or 2- hydroxy- ethoxyl methyl H or methyl C-4
##STR00069## H or 2- hydroxy- ethoxyl H or methyl H C-5
##STR00070## H or 2- hydroxy- ethoxyl H or methyl Methyl C-6
##STR00071## H or 2- hydroxy- ethoxyl H H C-7 ##STR00072## H or 2-
hydroxy- ethoxyl H methyl C-8 ##STR00073## H or 2- hydroxy- ethoxyl
methyl H C-9 ##STR00074## H or 2- hydroxy- ethoxyl methyl methyl
C-10 ##STR00075## H H or methyl H or methyl C-11 ##STR00076## H H H
or methyl C-12 ##STR00077## H methyl H or methyl C-13 ##STR00078##
H H or methyl H C-14 ##STR00079## H H or methyl methyl C-15
##STR00080## H H H C-16 ##STR00081## H methyl H C-17 ##STR00082## H
H methyl C-18 ##STR00083## H methyl methyl C-19 ##STR00084## 2-
hydroxy- ethoxyl H or methyl H or methyl C-20 ##STR00085## 2-
hydroxy- ethoxyl H H or methyl C-21 ##STR00086## 2-hydroxy- ethoxyl
methyl H or methyl C-22 ##STR00087## 2-hydroxy- ethoxyl H or methyl
H C-23 ##STR00088## 2-hydroxy- ethoxyl H or methyl methyl C-24
##STR00089## 2-hydroxy- ethoxyl H H C-25 ##STR00090## 2-hydroxy-
ethoxyl methyl H C-26 ##STR00091## 2-hydroxy- ethoxyl H methyl C-27
##STR00092## 2-hydroxy- ethoxyl methyl methyl
Separate embodiments herein, each providing a compound of Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof, in
which R.sup.1 is
##STR00093## include embodiments D-1 through D-27 wherein R.sup.2,
R.sup.3, and R.sup.4 are as defined in Table D for each of the
embodiments.
TABLE-US-00004 TABLE D Embodiment No. R.sup.1 R.sup.2 R.sup.3
R.sup.4 D-1 ##STR00094## H or 2- hydroxyethoxyl H or methyl H or
methyl D-2 ##STR00095## H or 2- hydroxyethoxyl H H or methyl D-3
##STR00096## H or 2- hydroxyethoxyl methyl H or methyl D-4
##STR00097## H or 2- hydroxyethoxyl H or methyl H D-5 ##STR00098##
H or 2- hydroxyethoxyl H or methyl Methyl D-6 ##STR00099## H or 2-
hydroxyethoxyl H H D-7 ##STR00100## H or 2- hydroxyethoxyl H methyl
D-8 ##STR00101## H or 2- hydroxyethoxyl methyl H D-9 ##STR00102## H
or 2- hydroxyethoxyl methyl methyl D-10 ##STR00103## H H or methyl
H or methyl D-11 ##STR00104## H H H or methyl D-12 ##STR00105## H
methyl H or methyl D-13 ##STR00106## H H or methyl H D-14
##STR00107## H H or methyl methyl D-15 ##STR00108## H H H D-16
##STR00109## H methyl H D-17 ##STR00110## H H methyl D-18
##STR00111## H methyl methyl D-19 ##STR00112## 2-hydroxyethoxyl H
or methyl H or methyl D-20 ##STR00113## 2-hydroxyethoxyl H H or
methyl D-21 ##STR00114## 2-hydroxyethoxyl methyl H or methyl D-22
##STR00115## 2-hydroxyethoxyl H or methyl H D-23 ##STR00116##
2-hydroxyethoxyl H or methyl methyl D-24 ##STR00117##
2-hydroxyethoxyl H H D-25 ##STR00118## 2-hydroxyethoxyl methyl H
D-26 ##STR00119## 2-hydroxyethoxyl H methyl D-27 ##STR00120##
2-hydroxyethoxyl methyl methyl
Embodiments herein that refer to the compound of Formula I or of
Formula II in one aspect also refer to a pharmaceutically
acceptable salt or co-crystal of the compound of Formula I or of
Formula II, even if not explicitly stated as such.
Also provided herein is a compound of Formula II:
##STR00121## or a pharmaceutically acceptable salt,
pharmaceutically acceptable co-crystal, pharmaceutically acceptable
ester, stereoisomer, mixture of stereoisomers or tautomer thereof,
wherein: R.sup.10 is selected from the group consisting of
##STR00122## wherein * indicates the carbon atom of the indicated
phenyl ring of Formula II to which R.sup.1 is attached; R.sup.20 is
H or 2-hydroxyethoxyl; R.sup.30 is H or methyl; and R.sup.40 is H,
halogen (i.e. F, Cl, Br, or I), methyl, or halo substituted methyl
(i.e. methyl wherein 1 to 3 hydrogen atoms are substituted by 1 to
3 halogen atoms, which may be the same or different, e.g.
fluoromethyl, chloromethyl, difluoromethyl, dichloromethyl,
chlorofluoromethyl, trifluoromethyl, and the like).
In some embodiments of the compound of Formula II, or a
pharmaceutically acceptable salt thereof, R.sup.10 is selected from
the group consisting of
##STR00123## In some embodiments, R.sup.10 is
##STR00124## In some embodiments, R.sup.10 is
##STR00125## In some embodiments, R.sup.10 is
##STR00126## Within each of the embodiments described herein
comprising a compound of Formula II, there is a further embodiment
wherein each of R.sup.20, R.sup.30, and R.sup.40 is H. Within each
of the embodiments described herein comprising a compound of
Formula II, there is a another embodiment wherein R.sup.20 is H,
R.sup.30 is methyl, and R.sup.40 is H. Within each of the
embodiments described herein comprising a compound of Formula II,
there is also another embodiment wherein R.sup.20 is H, R.sup.30 is
H, and R.sup.40 is methyl. Within each of the embodiments described
herein comprising a compound of Formula II, there is still another
embodiment wherein R.sup.20 is 2-hydroxyethoxyl, R.sup.30 is
methyl, and R.sup.40 is H. Within each of the embodiments described
herein comprising a compound of Formula II, there is still another
embodiment wherein R.sup.20 is 2-hydroxyethoxyl, R.sup.30 is
methyl, and R.sup.40 is H. Within each of the embodiments described
herein comprising a compound of Formula II, there is still further
embodiment wherein R.sup.20 is 2-hydroxyethoxyl, R.sup.30 is H, and
R.sup.40 is methyl.
Representative compounds of the invention are listed in Table A
below. The compounds in Table A are named using ChemBioDraw Ultra
12.0 and it should be understood that other names be used to
identify compounds of the same structure. Other compounds or
radicals may be named with common names, or systematic or
non-systematic names. The compounds may also be named using other
nomenclature systems and symbols that are commonly recognized in
the art of chemistry including, for example, Chemical Abstract
Service (CAS) and International Union of Pure and Applied Chemistry
(IUPAC). Any ambiguity in naming of compounds can be resolved by
deferring to the structure, where provided.
TABLE-US-00005 TABLE A Representative Compounds Structure Name
##STR00127## 6-(6-amino-5-methylpyrazin- 2-yl)-N-(4-(4-(oxetan-
3-yl)piperazin-1-yl)phenyl) imidazo[1,2-a]pyrazin-8-amine
##STR00128## 6-(6-aminopyrazin-2-yl)- N-(4-(4-(oxetan-3-
yl)piperazin-1-yl)phenyl) imidazo[1,2-a]pyrazin-8-amine
##STR00129## (R)-(4-(4-((6-(6-aminopyrazin- 2-yl)imidazo[1,2-
a]pyrazin-8-yl)amino)phenyl) morpholin-2-yl)methanol ##STR00130##
6-(6-aminopyrazin-2-yl)-5- methyl-N-(4-(4-(oxetan-
3-yl)piperazin-1-yl)phenyl) imidazo[1,2-a]pyrazin-8-amine
##STR00131## 2-(5-((6-(6-aminopyrazin- 2-yl)imidazo[1,2-
a]pyrazin-8-yl)amino)-2-(4- (oxetan-3-yl)piperazin-1-
yl)phenoxy)ethanol ##STR00132## 2-((4-(4-((6-(6-aminopyrazin-
2-yl)imidazo[1,2- a]pyrazin-8-yl)amino) phenyl)piperazin-1-
yl)methyl)propane-1,3-diol ##STR00133## 2-(5-((6-(6-amino-5-
methylpyrazin- 2-yl)imidazo[1,2- a]pyrazin-8-yl)amino)-2-
(4-(oxetan-3-yl)piperazin-1- yl)phenoxy)ethanol
An embodiment the invention provides
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine, pharmaceutically acceptable salts,
pharmaceutically acceptable co-crystals, pharmaceutically
acceptable esters, pharmaceutically acceptable solvates, hydrates,
enantiomers, mixture of enantiomers, tautomers, polymorphs, and
pharmaceutically acceptable prodrugs thereof. An embodiment of the
invention provides
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine or a pharmaceutically acceptable salt or
co-crystal thereof. An embodiment the invention provides
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine or a pharmaceutically acceptable salt
thereof.
The invention provides
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine mesylate, for example
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate. For example, the invention
provides
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form I, which may be
characterized by XRPD peaks at about 19.7, about 17.3, about 17.9,
about 21.6, and about 25.8 (2theta degrees). Also provided is
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form II, which may be
characterized by XRPD peaks at about 17.3, about 25.1, about 20.4,
about 19.6 and about 18.5 (2theta degrees).
An embodiment of the present invention provides
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine succinate. For example the invention
provides
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine succinate Form I, which may be
characterized by XRPD peaks at about 16.5, about 24.5, about 17.7,
about 28.4 and about 21.8 (2theta degrees). The invention also
provides
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine succinate Form II, which may be
characterized by XRPD peaks at about 25.0, about 16.3, about 22.0,
about 7.9, and about 7.6 (2theta degrees).
The term "about" as used in relation to XRPD peaks means, for
example, .+-.0.2, .+-.0.1, .+-.0.05 (2theta degrees) etc.
The compounds described herein, e.g. a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, or a
compound of Formula II, or a pharmaceutically acceptable salt or
co-crystal thereof, provide distinct advantages as Syk inhibitors.
The compounds described herein are inhibitors of Syk kinase
activity, as measured, for example, as the inhibition of Syk kinase
activity in a biochemical assay or as the reduction in basophil
activation as measured by CD63 expression, as described in the
Examples. The compounds described herein also have desirable
properties for use as a pharmaceutical, including kinetic
solubility at 37.degree. C. in phosphate buffer at pH 7.4 and low
levels of hepatocyte clearance. These features result in Syk
inhibitors for treatment of disease with pharmacokinetic
characteristics that provide a therapeutic window such that the
compounds can be effective in smaller doses than currently known
compounds. As such, the compounds provide effective doses with
minimal off target activity, which may reduce unwanted side
effects, lessen the chance of drug-drug interactions, and increase
a subject's compliance with a given therapeutic regimen.
In some embodiments, the compounds described herein, e.g. a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, or a compound of Formula II, or a
pharmaceutically acceptable salt or co-crystal thereof, is
effective in one or more of Syk kinase activity inhibition or
reduction of basophil activation as measured by CD63 expression,
for example, the compound inhibits Syk kinase activity with an
IC.sub.50 value less than or equal to 1 micromolar, less than or
equal to 500 nanomolar, less than or equal to 200 nanomolar, less
than or equal to 100 nanomolar, less than or equal to 50 nanomolar,
less than or equal to 20 nanomolar, or less than or equal to 10
nanomolar, as demonstrated by a suitable assay for Syk kinase
activity, such as the assay as described in Example 12; and/or
reduces CD63 expression activity with an EC.sub.50 value less than
or equal to 1 micromolar, less than or equal to 500 nanomolar, less
than or equal to 200 nanomolar, less than or equal to 150
nanomolar, less than or equal to 100 nanomolar, or less than or
equal to 75 nanomolar, as demonstrated by a suitable assay for the
measurement of CD63 expression in basophils, such as the assay as
described in Example 9.
In some embodiments, the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, is
effective in both of Syk kinase inhibition and reduction of CD63
expression, for example, the compound has Syk kinase activity with
an IC.sub.50 value less than or equal to 1 micromolar, less than or
equal to 500 nanomolar, less than or equal to 200 nanomolar, less
than or equal to 100 nanomolar, less than or equal to 50 nanomolar,
less than or equal to 20 nanomolar, or less than or equal to 10
nanomolar, as demonstrated by a suitable assay for Syk kinase
activity, such as the assay as described in Example 12; and has
reduction in CD63 expression with an EC.sub.50 value less than or
equal to 1 micromolar, less than or equal to 500 nanomolar, less
than or equal to 200 nanomolar, less than or equal to 150
nanomolar, less than or equal to 100 nanomolar, or less than or
equal to 75 nanomolar, as demonstrated by a suitable assay for the
measurement of CD63 expression in basophils, such as the assay as
described in Example 9.
In some embodiments, in addition to having the property of one or
more of Syk kinase inhibition or reduction of basophil activation
as measured by CD63 expression, including having both of the
properties of Syk kinase inhibition and reduction of basophil
activation as measured by CD63 expression, the compound of Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof, has
desirable properties for use as a pharmaceutical, including one or
more of kinetic solubility and low levels of hepatocyte clearance.
In some embodiments, the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, has a
desirable property of one or more of kinetic solubility and low
levels of hepatocyte clearance, including kinetic solubility at
37.degree. C. in phosphate buffer at pH 7.4 of greater than or
equal to 10 .mu.M, greater than or equal to 20 .mu.M, greater than
or equal to 30 .mu.M, greater than or equal to 40 .mu.M, greater
than or equal to 50 .mu.M, greater than or equal to 60 .mu.M,
greater than or equal to 70 .mu.M, greater than or equal to 80
.mu.M, or greater than or equal to 90 .mu.M, as demonstrated by a
suitable measure of kinetic solubility, such as the assay as
described in Example 10; and/or predicted hepatocyte clearance of
less than or equal to 0.50 L/hr/kg, less than or equal to 0.40
L/hr/kg, less than or equal to 0.30 L/hr/kg, less than or equal to
0.20 L/hr/kg, less than or equal to 0.10 L/hr/kg, less than or
equal to 0.09 L/hr/kg, less than or equal to 0.08 L/hr/kg, less
than or equal to 0.07 L/hr/kg, or less than or equal to 0.06
L/hr/kg, as demonstrated by a suitable measure of predicted
hepatocyte clearance, such as the assay as described in Example
11.
In some embodiments, the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, has a
desirable property of kinetic solubility, and low levels of
hepatocyte clearance, including kinetic solubility at 37.degree. C.
in phosphate buffer at pH 7.4 of greater than or equal to 10 .mu.M,
greater than or equal to 20 .mu.M, greater than or equal to 30
.mu.M, greater than or equal to 40 .mu.M, greater than or equal to
50 .mu.M, greater than or equal to 60 .mu.M, greater than or equal
to 70 .mu.M, greater than or equal to 80 .mu.M, or greater than or
equal to 90 .mu.M, as demonstrated by a suitable measure of kinetic
solubility, such as the assay as described in Example 10; and
predicted hepatocyte clearance of less than or equal to 0.50
L/hr/kg, less than or equal to 0.40 L/hr/kg, less than or equal to
0.30 L/hr/kg, less than or equal to 0.20 L/hr/kg, less than or
equal to 0.10 L/hr/kg, less than or equal to 0.09 L/hr/kg, less
than or equal to 0.08 L/hr/kg, less than or equal to 0.07 L/hr/kg,
or less than or equal to 0.06 L/hr/kg, as demonstrated by a
suitable measure of predicted hepatocyte clearance, such as the
assay as described in Example 11.
In some embodiments, the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, is
effective in both of Syk kinase inhibition and reduction of CD63
expression, and has a desirable property of kinetic solubility, and
low levels of hepatocyte clearance, for example, the compound has
Syk kinase activity with an IC.sub.50 value less than or equal to 1
micromolar, less than or equal to 500 nanomolar, less than or equal
to 200 nanomolar, less than or equal to 100 nanomolar, less than or
equal to 50 nanomolar, less than or equal to 20 nanomolar, or less
than or equal to 10 nanomolar, as demonstrated by a suitable assay
for Syk kinase activity, such as the assay as described in Example
12; and has reduced CD63 expression with an EC.sub.50 value less
than or equal to 1 micromolar, less than or equal to 500 nanomolar,
less than or equal to 200 nanomolar, less than or equal to 150
nanomolar, less than or equal to 100 nanomolar, or less than or
equal to 75 nanomolar, as demonstrated by a suitable assay for the
measurement of CD63 expression in basophils, such as the assay as
described in Example 10; and kinetic solubility at 37.degree. C. in
phosphate buffer at pH 7.4 of greater than or equal to 10 .mu.M,
greater than or equal to 20 .mu.M, greater than or equal to 30
.mu.M, greater than or equal to 40 .mu.M, greater than or equal to
50 .mu.M, greater than or equal to 60 .mu.M, greater than or equal
to 70 .mu.M, greater than or equal to 80 .mu.M, or greater than or
equal to 90 .mu.M, as demonstrated by a suitable measure of kinetic
solubility, such as the assay as described in Example 10; and
predicted hepatocyte clearance of less than or equal to 0.50
L/hr/kg, less than or equal to 0.40 L/hr/kg, less than or equal to
0.30 L/hr/kg, less than or equal to 0.20 L/hr/kg, less than or
equal to 0.10 L/hr/kg, less than or equal to 0.09 L/hr/kg, less
than or equal to 0.08 L/hr/kg, less than or equal to 0.07 L/hr/kg,
or less than or equal to 0.06 L/hr/kg, as demonstrated by a
suitable measure of predicted hepatocyte clearance, such as the
assay as described in Example 11.
Methods of Use
The invention provides a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, for use in
therapy. Provided is a method of treating a subject, for example, a
mammal, such as a human, having a disease responsive to inhibition
of Syk activity, comprising administrating to the subject having,
or suspected of having, such a disease, an effective amount of a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof. In one aspect, the subject, such as a human, is
administered a pharmaceutical composition comprising a compound of
Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, and a pharmaceutically acceptable vehicle. The invention
further provides a compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof for use in such methods.
In some embodiments, the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, may be
administered to a subject (e.g., a human) who is at risk or has a
family history of the disease or condition.
In some embodiments, the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, may also
inhibit other kinases, such that disease, disease symptoms, and
conditions associated with these kinases are also treated.
Methods of treatment also include inhibiting Syk activity and/or
inhibiting B-cell activity, by inhibiting ATP binding or hydrolysis
by Syk or by some other mechanism, in vivo, in a subject suffering
from a disease responsive to inhibition of Syk activity, by
administering an effective concentration of a compound of Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof. An
example of an effective concentration would be that concentration
sufficient to inhibit Syk activity in vitro. An effective
concentration may be ascertained experimentally, for example by
assaying blood concentration of the compound following
administration to a human, or theoretically, by calculating
bioavailability.
In some embodiments, the condition responsive to inhibition of Syk
activity and/or B-cell activity is cancer, an allergic disorder
and/or an autoimmune and/or inflammatory disease, and/or an acute
inflammatory reaction.
Also provided is a method of inhibiting B-cell activity in a
subject in need thereof comprising administering an effective
amount of a compound of Formula I, or a pharmaceutically acceptable
salt or co-crystal thereof.
Also provided is a method of inhibiting B-cell proliferation in a
subject in need thereof comprising administering an effective
amount of a compound of Formula I, or a pharmaceutically acceptable
salt or co-crystal thereof.
Also provided is a method of treating a subject having cancer, an
allergic disorder and/or an autoimmune and/or inflammatory disease,
and/or an acute inflammatory reaction, by administering an
effective amount of a compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof.
In some embodiments, the conditions and diseases that can be
treated using a compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, include, but are not limited
tolymphoma (e.g. small lymphocytic lymphoma (SLL), non-Hodgkin's
lymphoma (NHL), indolent non-Hodgkin's lymphoma (iNHL), refractory
iNHL, mantle cell lymphoma (MCL), follicular lymphoma (FL),
lymphoplasmacytic lymphoma (LPL), marginal zone lymphoma (MZL),
immunoblastic large cell lymphoma, lymphoblastic lymphoma, Splenic
marginal zone B-cell lymphoma (+/-villous lymphocytes), Nodal
marginal zone lymphoma (+/-monocytoid B-cells), Extranodal marginal
zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT)
type, T-cell lymphoma (e.g. cutaneous T-cell lymphoma, extranodal
T-cell lymphoma, anaplastic large cell lymphoma, angioimmunoblastic
T-cell lymphoma, mycosis fungoides), B-cell lymphoma, diffuse large
B-cell lymphoma (DLBCL), Mediastinal large B-cell lymphoma,
Intravascular large B-cell lymphoma, Primary effusion lymphoma,
small non-cleaved cell lymphoma, or Burkitt's lymphoma), multiple
myeloma, plasmacytoma, and leukemia (e.g. acute lymphocytic
leukemia (ALL), T-cell acute lymphoblastic leukemia (T-ALL), B-cell
acute lymphoblastic leukemia (B-ALL), B-cell prolymphocytic
leukemia, acute myeloid leukemia (AML), chronic lymphocytic
leukemia (CLL), juvenile myelomonocytic leukemia (JMML), minimal
residual disease (MRD), hairy cell leukemia, myelofibrosis (e.g.
primary or secondary myelofibrosis), or chronic myeloid leukemia
(CML), myelodysplastic syndrome (MDS), myeloproliferative disease
(MPD), Waldestrom's macroglobulinemia (WM), polycythemia vera,
essential thrombocythemia, pancreatic cancer, urological cancer,
bladder cancer, colorectal cancer, colon cancer, breast cancer,
prostate cancer, renal cancer, hepatocellular cancer, thyroid
cancer, gall bladder cancer, lung cancer (e.g. non-small cell lung
cancer, small-cell lung cancer), ovarian cancer, cervical cancer,
gastric cancer, endometrial cancer, esophageal cancer, head and
neck cancer, melanoma, neuroendocrine cancer, CNS cancer, brain
tumors (e.g., glioma, anaplastic oligodendroglioma, adult
glioblastoma multiforme, and adult anaplastic astrocytoma), bone
cancer, soft tissue sarcoma, retinoblastomas, neuroblastomas,
peritoneal effusions, malignant pleural effusions, mesotheliomas,
Wilms tumors, trophoblastic neoplasms, hemangiopericytomas,
Kaposi's sarcomas, myxoid carcinoma, round cell carcinoma, squamous
cell carcinomas, esophageal squamous cell carcinomas, oral
carcinomas, cancers of the adrenal cortex, ACTH-producing tumors,
systemic lupus erythematosus (SLE), myestenia gravis, Goodpasture's
syndrome, glomerulonephritis, hemorrhage, pulmonary hemorrhage,
atherosclerosis, rheumatoid arthritis (RA), psoriatic arthritis,
monoarticular arthritis, osteoarthritis, gouty arthritis,
spondylitis, Behcet disease, autoimmune thyroiditis, Reynaud's
syndrome, acute disseminated encephalomyelitis, chronic idiopathic
thrombocytopenic purpura, multiple sclerosis (MS), Sjogren's
syndrome, autoimmune hemolytic anemia, tissue graft rejection,
hyperacute rejection of transplanted organs, allograft rejection,
graft-versus-host disease, diseases involving leukocyte diapedesis,
disease states due to leukocyte dyscrasia and metastasis,
granulocyte transfusion-associated syndromes, cytokine-induced
toxicity, scleroderma, vasculitis, asthma, psoriasis, inflammatory
bowel disease (e.g. chronic inflammatory bowel disease, ulcerative
colitis, Crohn's disease, necrotizing enterocolitis), irritable
bowel syndrome, dermatomyositis, Addison's disease, Parkinson's
disease, Alzheimer's disease, diabetes, type I diabetes mellitus,
sepsis, septic shock, endotoxic shock, gram negative sepsis, gram
positive sepsis, and toxic shock syndrome, multiple organ injury
syndrome secondary to septicemia, trauma, hypovolemic shock,
allergic conjunctivitis, vernal conjunctivitis, and
thyroid-associated ophthalmopathy, eosinophilic granuloma, eczema,
chronic bronchitis, acute respiratory distress syndrome, allergic
rhinitis, coryza, hay fever, bronchial asthma, silicosis, pulmonary
sarcoidosis, pleurisy, alveolitis, emphysema, pneumonia, bacterial
pneumonia, bronchiectasis, and pulmonary oxygen toxicity,
reperfusion injury of the myocardium, brain, or extremities,
thermal injury, cystic fibrosis, keloid formation or scar tissue
formation, fever and myalgias due to infection, and brain or spinal
cord injury due to minor trauma, diseases involving leukocyte
diapedesis, acute hypersensitivity, delayed hypersensitivity,
urticaria, food allergies, skin sunburn, inflammatory pelvic
disease, urethritis, uveitis, sinusitis, pneumonitis, encephalitis,
meningitis, myocarditis, nephritis, osteomyelitis, myositis,
hepatitis, alcoholic hepatitis, gastritis, enteritis, contact
dermatitis, atopic dermatitis, gingivitis, appendicitis,
pancreatitis, cholocystitis, and polycystic kidney disease.
In some embodiments, provided is a method of treating a subject
having an allergic disorder and/or an autoimmune and/or
inflammatory disease, and/or an acute inflammatory reaction by
administering an effective amount of a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof. In some
embodiments, the disease is selected from the group consisting of
systemic lupus erythematosus, myestenia gravis, Goodpasture's
syndrome, glomerulonephritis, hemorrhage, pulmonary hemorrhage,
atherosclerosis, rheumatoid arthritis, psoriatic arthritis,
monoarticular arthritis, osteoarthritis, gouty arthritis,
spondylitis, Behcet disease, autoimmune thyroiditis, Reynaud's
syndrome, acute disseminated encephalomyelitis, chronic idiopathic
thrombocytopenic purpura, multiple sclerosis, Sjogren's syndrome,
autoimmune hemolytic anemia, tissue graft rejection, hyperacute
rejection of transplanted organs, allograft rejection,
graft-versus-host disease, diseases involving leukocyte diapedesis,
disease states due to leukocyte dyscrasia and metastasis,
granulocyte transfusion-associated syndromes, cytokine-induced
toxicity, scleroderma, vasculitis, asthma, psoriasis, chronic
inflammatory bowel disease, ulcerative colitis, Crohn's disease,
necrotizing enterocolitis, irritable bowel syndrome,
dermatomyositis, Addison's disease, Parkinson's disease,
Alzheimer's disease, diabetes, type I diabetes mellitus, sepsis,
septic shock, endotoxic shock, gram negative sepsis, gram positive
sepsis, and toxic shock syndrome, multiple organ injury syndrome
secondary to septicemia, trauma, hypovolemic shock, allergic
conjunctivitis, vernal conjunctivitis, and thyroid-associated
ophthalmopathy, eosinophilic granuloma, eczema, chronic bronchitis,
acute respiratory distress syndrome, allergic rhinitis, coryza, hay
fever, bronchial asthma, silicosis, pulmonary sarcoidosis,
pleurisy, alveolitis, emphysema, pneumonia, bacterial pneumonia,
bronchiectasis, and pulmonary oxygen toxicity, reperfusion injury
of the myocardium, brain, or extremities, thermal injury, cystic
fibrosis, keloid formation or scar tissue formation, fever and
myalgias due to infection, and brain or spinal cord injury due to
minor trauma, diseases involving leukocyte diapedesis, acute
hypersensitivity, delayed hypersensitivity, urticaria, food
allergies, skin sunburn, inflammatory pelvic disease, urethritis,
uveitis, sinusitis, pneumonitis, encephalitis, meningitis,
myocarditis, nephritis, osteomyelitis, myositis, hepatitis,
alcoholic hepatitis, gastritis, enteritis, contact dermatitis,
atopic dermatitis, gingivitis, appendicitis, pancreatitis,
cholocystitis, and polycystic kidney disease.
In some embodiments, provided is a method of treating a subject
having an autoimmune disease selected from the group consisting of
a systemic lupus erythematosus, myestenia gravis, rheumatoid
arthritis, acute disseminated encephalomyelitis, idiopathic
thrombocytopenic purpura, multiple sclerosis, Sjoegren's syndrome,
psoriasis, autoimmune hemolytic anemia, asthma, ulcerative colitis,
Crohn's disease, irritable bowel disease, and chronic obstructive
pulmonary disease by administering an effective amount of a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof. In some embodiments, the autoimmune disease has
excessive or destructive immune reactions, such as asthma,
rheumatoid arthritis, multiple sclerosis, chronic obstructive
pulmonary disease, or systemic lupus erythematosus.
In some embodiments, provided is a method of treating a subject
having rheumatoid arthritis, by administering an effective amount
of a compound of Formula I, or a pharmaceutically acceptable salt
or co-crystal thereof.
Syk is a known inhibitor of apoptosis in lymphoma B-cells.
Defective apoptosis contributes to the pathogenesis and drug
resistance of human leukemias and lymphomas. Thus, further provided
is a method of promoting or inducing apoptosis in cells expressing
Syk comprising contacting the cell with a compound of Formula I, or
a pharmaceutically acceptable salt or co-crystal thereof.
In some embodiments, provided is a method of treating a subject
having cancer selected from the group consisting of carcinoma,
sarcoma, melanoma, lymphoma and leukemia. In some embodiments the
cancer is a solid tumor or a hematologic malignancy.
In some embodiments, provided is a method of treating a subject
having a hematologic malignancy selected from the group consisting
of small lymphocytic lymphoma, non-Hodgkin's lymphoma, indolent
non-Hodgkin's lymphoma, refractory iNHL, mantle cell lymphoma,
follicular lymphoma, lymphoplasmacytic lymphoma, marginal zone
lymphoma, immunoblastic large cell lymphoma, lymphoblastic
lymphoma, Splenic marginal zone B-cell lymphoma (+/-villous
lymphocytes), Nodal marginal zone lymphoma (+/-monocytoid B-cells),
Extranodal marginal zone B-cell lymphoma of mucosa-associated
lymphoid tissue type, cutaneous T-cell lymphoma, extranodal T-cell
lymphoma, anaplastic large cell lymphoma, angioimmunoblastic T-cell
lymphoma, mycosis fungoides, B-cell lymphoma, diffuse large B-cell
lymphoma, Mediastinal large B-cell lymphoma, Intravascular large
B-cell lymphoma, Primary effusion lymphoma, small non-cleaved cell
lymphoma, Burkitt's lymphoma, multiple myeloma, plasmacytoma, acute
lymphocytic leukemia, T-cell acute lymphoblastic leukemia, B-cell
acute lymphoblastic leukemia, B-cell prolymphocytic leukemia, acute
myeloid leukemia, chronic lymphocytic leukemia, juvenile
myelomonocytic leukemia, minimal residual disease, hairy cell
leukemia, primary myelofibrosis, secondary myelofibrosis, chronic
myeloid leukemia, myelodysplastic syndrome, myeloproliferative
disease, and Waldestrom's macroglobulinemia.
In some embodiments, provided is a method of treating a subject
having cancer, wherein the cancer is leukemia or lymphoma, by
administering an effective amount of a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof. In some
embodiments, the cancer is selected from the group consisting of
acute lymphocytic leukemia, acute myeloid leukemia, chronic
lymphocytic leukemia, small lymphocytic lymphoma, myelodysplastic
syndrome, myeloproliferative disease, chronic myeloid leukemia,
multiple myeloma, indolent non-Hodgkin's lymphoma, refractory iNHL,
non-Hodgkin's lymphoma, mantle cell lymphoma, follicular lymphoma,
Waldestrom's macroglobulinemia, T-cell lymphoma, B-cell lymphoma,
and diffuse large B-cell lymphoma. In one embodiment, the cancer is
T-cell acute lymphoblastic leukemia, or B-cell acute lymphoblastic
leukemia. The non-Hodgkin lymphoma encompasses the indolent B-cell
diseases that include, for example, follicular lymphoma,
lymphoplasmacytic lymphoma, Waldenstrom macroglobulinemia, and
marginal zone lymphoma, as well as the aggressive lymphomas that
include, for example, Burkitt's lymphoma, diffuse large B-cell
lymphoma and mantle cell lymphoma. In one embodiment, the cancer is
indolent non-Hodgkin's lymphoma.
In some embodiments, provided is a method of treating a subject
having a hematologic malignancy by administering an effective
amount of a compound of Formula I, or a pharmaceutically acceptable
salt or co-crystal thereof. In specific embodiments, the
hematologic malignancy is leukemia (e.g., chronic lymphocytic
leukemia) or lymphoma (e.g., non-Hodgkin's lymphoma).
In some embodiments, provided is a method of treating a subject
having chronic lymphocytic leukemia by administering an effective
amount of a compound of Formula I, or a pharmaceutically acceptable
salt or co-crystal thereof.
In some embodiments, provided is a method of treating a subject
having a solid tumor by administering an effective amount of a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof. In some embodiments, the solid tumor is from a
cancer selected from the group consisting of pancreatic cancer,
urological cancer, bladder cancer, colorectal cancer, colon cancer,
breast cancer, prostate cancer, renal cancer, hepatocellular
cancer, thyroid cancer, gall bladder cancer, lung cancer (e.g.
non-small cell lung cancer, small-cell lung cancer), ovarian
cancer, cervical cancer, gastric cancer, endometrial cancer,
esophageal cancer, head and neck cancer, melanoma, neuroendocrine
cancer, CNS cancer, brain tumors (e.g., glioma, anaplastic
oligodendroglioma, adult glioblastoma multiforme, and adult
anaplastic astrocytoma), bone cancer, soft tissue sarcoma,
retinoblastomas, neuroblastomas, peritoneal effusions, malignant
pleural effusions, mesotheliomas, Wilms tumors, trophoblastic
neoplasms, hemangiopericytomas, Kaposi's sarcomas, myxoid
carcinoma, round cell carcinoma, squamous cell carcinomas,
esophageal squamous cell carcinomas, oral carcinomas, cancers of
the adrenal cortex, and ACTH-producing tumors. In some embodiments,
the solid tumor is from non-small cell lung cancer, small-cell lung
cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal
cancer, prostate cancer, and breast cancer.
Also provided herein is a compound as described herein, e.g. a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, for use in the treatment of a disease or
condition as described herein, e.g. a cancer (including carcinoma,
sarcoma, melanoma, lymphoma and leukemia), an allergic disorder
and/or an autoimmune and/or inflammatory disease, and/or an acute
inflammatory reaction. Also provided herein is a compound of
formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, for use in a method of treatment described in this
disclosure.
Also provided herein is a compound as described herein, e.g. a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, for use in the manufacture of a medicament for
the treatment of a disease or condition as described herein, e.g. a
cancer (including carcinoma, sarcoma, melanoma, lymphoma and
leukemia), an allergic disorder and/or an autoimmune and/or
inflammatory disease, and/or an acute inflammatory reaction.
Subjects
Any of the methods of treatment provided may be used to treat a
subject who has been diagnosed with or is suspected of having an
allergic disorder and/or an autoimmune and/or inflammatory disease,
and/or an acute inflammatory reaction or a cancer.
In some of the embodiments of any of the methods provided herein,
the subject is a human who is at risk of developing a cancer (e.g.,
a human who is genetically or otherwise predisposed to developing a
cancer) and who has or has not been diagnosed with the cancer. As
used herein, an "at risk" subject is a subject who is at risk of
developing cancer (e.g., a hematologic malignancy). The subject may
or may not have detectable disease, and may or may not have
displayed detectable disease prior to the treatment methods
described herein. An at risk subject may have one or more so-called
risk factors, which are measurable parameters that correlate with
development of cancer, such as described herein. A subject having
one or more of these risk factors has a higher probability of
developing cancer than an individual without these risk
factor(s).
These risk factors may include, for example, age, sex, race, diet,
history of previous disease, presence of precursor disease, genetic
(e.g., hereditary) considerations, and environmental exposure. In
some embodiments, a subject at risk for cancer includes, for
example, a subject whose relatives have experienced this disease,
and those whose risk is determined by analysis of genetic or
biochemical markers. Prior history of having cancer may also be a
risk factor for instances of cancer recurrence.
Provided herein are also methods for treating a subject (e.g., a
human) who exhibits one or more symptoms associated with cancer
(e.g., a hematologic malignancy). In some embodiments, the subject
is at an early stage of cancer. In other embodiments, the subject
is at an advanced stage of cancer.
Provided herein are also methods for treating a subject (e.g., a
human) who is undergoing one or more standard therapies for
treating cancer (e.g., a hematologic malignancy), such as
chemotherapy, radiotherapy, immunotherapy, and/or surgery. Thus, in
some foregoing embodiments, the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof,
administered before, during, or after administration of
chemotherapy, radiotherapy, immunotherapy, and/or surgery.
In another aspect, provided herein are methods for treating a
subject (e.g., a human) who is "refractory" to a cancer treatment
or who is in "relapse" after treatment for cancer (e.g., a
hematologic malignancy). A subject "refractory" to an anti-cancer
therapy means they do not respond to the particular treatment, also
referred to as resistant. The cancer may be resistant to treatment
from the beginning of treatment, or may become resistant during the
course of treatment, for example after the treatment has shown some
effect on the cancer, but not enough to be considered a remission
or partial remission. A subject in "relapse" means that the cancer
has returned or the signs and symptoms of cancer have returned
after a period of improvement, e.g. after a treatment has shown
effective reduction in the cancer, such as after a subject is in
remission or partial remission.
In some embodiments, the subject may be a human who is (i)
refractory to at least one anti-cancer therapy, or (ii) in relapse
after treatment with at least one anti-cancer therapy, or both (i)
and (ii). In some of embodiments, the subject is refractory to at
least two, at least three, or at least four anti-cancer therapies
(including, for example, standard or experimental
chemotherapies).
In some embodiments, the subject is refractory to at least one, at
least two, at least three, or at least four anti-cancer therapies
(including, for example, standard or experimental chemotherapy)
selected from fludarabine, rituximab, obinutuzumab, alkylating
agents, alemtuzumab and other chemotherapy treatments such as CHOP
(cyclophosphamide, doxorubicin, vincristine, prednisone); R-CHOP
(rituximab-CHOP); hyperCVAD (hyperfractionated cyclophosphamide,
vincristine, doxorubicin, dexamethasone, methotrexate, cytarabine);
R-hyperCVAD (rituximab-hyperCVAD); FCM (fludarabine,
cyclophosphamide, mitoxantrone); R-FCM (rituximab, fludarabine,
cyclophosphamide, mitoxantrone); bortezomib and rituximab;
temsirolimus and rituximab; temsirolimus and Velcade.RTM.;
Iodine-131 tositumomab (Bexxar.RTM.) and CHOP; CVP
(cyclophosphamide, vincristine, prednisone); R-CVP (rituximab-CVP);
ICE (iphosphamide, carboplatin, etoposide); R-ICE (rituximab-ICE);
FCR (fludarabine, cyclophosphamide, rituximab); FR (fludarabine,
rituximab); D.T. PACE (dexamethasone, thalidomide, cisplatin,
Adriamycin.RTM., cyclophosphamide, etoposide); and idelalisib.
Other examples of chemotherapy treatments (including standard or
experimental chemotherapies) are described below. In addition,
treatment of certain lymphomas is reviewed in Cheson, B. D.,
Leonard, J. P., "Monoclonal Antibody Therapy for B-Cell
Non-Hodgkin's Lymphoma" The New England Journal of Medicine 2008,
359(6), p. 613-626; and Wierda, W. G., "Current and Investigational
Therapies for Patients with CLL" Hematology 2006, p. 285-294.
Lymphoma incidence patterns in the United States is profiled in
Morton, L. M., et al. "Lymphoma Incidence Patterns by WHO Subtype
in the United States, 1992-2001" Blood 2006, 107(1), p.
265-276.
For example, treatment of non-Hodgkin's lymphomas (NHL), especially
of B-cell origin, include the use of monoclonal antibodies,
standard chemotherapy approaches (e.g., CHOP, CVP, FCM, MCP, and
the like), radioimmunotherapy, and combinations thereof, especially
integration of an antibody therapy with chemotherapy. Examples of
unconjugated monoclonal antibodies for Non-Hodgkin's
lymphoma/B-cell cancers include rituximab, alemtuzumab, human or
humanized anti-CD20 antibodies, lumiliximab, anti-TRAIL,
bevacizumab, galiximab, epratuzumab, SGN-40, and anti-CD74.
Examples of experimental antibody agents used in treatment of
Non-Hodgkin's lymphoma/B-cell cancers include ofatumumab, ha20,
PRO131921, alemtuzumab, galiximab, SGN-40, CHIR-12.12, epratuzumab,
lumiliximab, apolizumab, milatuzumab, and bevacizumab. Examples of
standard regimens of chemotherapy for Non-Hodgkin's lymphoma/B-cell
cancers include CHOP (cyclophosphamide, doxorubicin, vincristine,
prednisone), FCM (fludarabine, cyclophosphamide, mitoxantrone), CVP
(cyclophosphamide, vincristine and prednisone), MCP (mitoxantrone,
chlorambucil, and prednisolone), R-CHOP (rituximab plus CHOP),
R-FCM (rituximab plus FCM), R-CVP (rituximab plus CVP), and R-MCP
(R-MCP). Examples of radioimmunotherapy for Non-Hodgkin's
lymphoma/B-cell cancers include yttrium-90-labeled ibritumomab
tiuxetan, and iodine-131-labeled tositumomab.
In another example, therapeutic treatments for mantle cell lymphoma
(MCL) include combination chemotherapies such as CHOP
(cyclophosphamide, doxorubicin, vincristine, prednisone), hyperCVAD
(hyperfractionated cyclophosphamide, vincristine, doxorubicin,
dexamethasone, methotrexate, cytarabine) and FCM (fludarabine,
cyclophosphamide, mitoxantrone). In addition, these regimens can be
supplemented with the monoclonal antibody rituximab (Rituxan) to
form combination therapies R-CHOP, hyperCVAD-R, and R-FCM. Other
approaches include combining any of the abovementioned therapies
with stem cell transplantation or treatment with ICE (iphosphamide,
carboplatin and etoposide). Other approaches to treating mantle
cell lymphoma includes immunotherapy such as using monoclonal
antibodies like Rituximab (Rituxan). Rituximab can be used for
treating indolent B-cell cancers, including marginal-zone lymphoma,
WM, CLL and small lymphocytic lymphoma. A combination of Rituximab
and chemotherapy agents is especially effective. A modified
approach is radioimmunotherapy, wherein a monoclonal antibody is
combined with a radioisotope particle, such as Iodine-131
tositumomab (Bexxar.RTM.) and Yttrium-90 ibritumomab tiuxetan
(Zevalin.RTM.). In another example, Bexxar.RTM. is used in
sequential treatment with CHOP. Another immunotherapy example
includes using cancer vaccines, which is based upon the genetic
makeup of an individual subject's tumor. A lymphoma vaccine example
is GTOP-99 (MyVax.RTM.). Yet other approaches to treating mantle
cell lymphoma includes autologous stem cell transplantation coupled
with high-dose chemotherapy, or treating mantle cell lymphoma
includes administering proteasome inhibitors, such as Velcade.RTM.
(bortezomib or PS-341), or antiangiogenesis agents, such as
thalidomide, especially in combination with Rituxan. Another
treatment approach is administering drugs that lead to the
degradation of Bcl-2 protein and increase cancer cell sensitivity
to chemotherapy, such as oblimersen (Genasense) in combination with
other chemotherapeutic agents. Another treatment approach includes
administering mTOR inhibitors, which can lead to inhibition of cell
growth and even cell death; a non-limiting example is Temsirolimus
(CCI-779), and Temsirolimus in combination with Rituxan.RTM.,
Velcade.RTM. or other chemotherapeutic agents.
Other recent therapies for MCL have been disclosed (Nature Reviews;
Jares, P. 2007). Such examples include Flavopiridol, PD0332991,
R-roscovitine (Selicilib, CYC202), Styryl sulphones, Obatoclax
(GX15-070), TRAIL, Anti-TRAIL DR4 and DR5 antibodies, Temsirolimus
(CCl-779), Everolimus (RAD001), BMS-345541, Curcumin, Vorinostat
(SAHA), Thalidomide, lenalidomide (Revlimid.RTM., CC-5013), and
Geldanamycin (17-AAG).
Examples of other therapeutic agents used to treat Waldenstrom's
Macroglobulinemia (WM) include perifosine, bortezomib
(Velcade.RTM.), rituximab, sildenafil citrate (Viagra.RTM.),
CC-5103, thalidomide, epratuzumab (hLL2-anti-CD22 humanized
antibody), simvastatin, enzastaurin, campath-1H, dexamethasone, DT
PACE, oblimersen, antineoplaston A10, antineoplaston AS2-1,
alemtuzumab, beta alethine, cyclophosphamide, doxorubicin
hydrochloride, prednisone, vincristine sulfate, fludarabine,
filgrastim, melphalan, recombinant interferon alfa, carmustine,
cisplatin, cyclophosphamide, cytarabine, etoposide, melphalan,
dolastatin 10, indium In 111 monoclonal antibody MN-14, yttrium Y
90 humanized epratuzumab, anti-thymocyte globulin, busulfan,
cyclosporine, methotrexate, mycophenolate mofetil, therapeutic
allogeneic lymphocytes, Yttrium Y 90 ibritumomab tiuxetan,
sirolimus, tacrolimus, carboplatin, thiotepa, paclitaxel,
aldesleukin, recombinant interferon alfa, docetaxel, ifosfamide,
mesna, recombinant interleukin-12, recombinant interleukin-11,
Bcl-2 family protein inhibitor ABT-263, denileukin diftitox,
tanespimycin, everolimus, pegfilgrastim, vorinostat, alvocidib,
recombinant flt3 ligand, recombinant human thrombopoietin,
lymphokine-activated killer cells, amifostine trihydrate,
aminocamptothecin, irinotecan hydrochloride, caspofungin acetate,
clofarabine, epoetin alfa, nelarabine, pentostatin, sargramostim,
vinorelbine ditartrate, WT-1 analog peptide vaccine, WT1 126-134
peptide vaccine, fenretinide, ixabepilone, oxaliplatin, monoclonal
antibody CD19, monoclonal antibody CD20, omega-3 fatty acids,
mitoxantrone hydrochloride, octreotide acetate, tositumomab and
iodine I-131 tositumomab, motexafin gadolinium, arsenic trioxide,
tipifarnib, autologous human tumor-derived HSPPC-96, veltuzumab,
bryostatin 1, and PEGylated liposomal doxorubicin hydrochloride,
and any combination thereof.
Examples of therapeutic procedures used to treat WM include
peripheral blood stem cell transplantation, autologous
hematopoietic stem cell transplantation, autologous bone marrow
transplantation, antibody therapy, biological therapy, enzyme
inhibitor therapy, total body irradiation, infusion of stem cells,
bone marrow ablation with stem cell support, in vitro-treated
peripheral blood stem cell transplantation, umbilical cord blood
transplantation, immunoenzyme technique, pharmacological study,
low-LET cobalt-60 gamma ray therapy, bleomycin, conventional
surgery, radiation therapy, and nonmyeloablative allogeneic
hematopoietic stem cell transplantation.
Examples of other therapeutic agents used to treat diffuse large
B-cell lymphoma (DLBCL) drug therapies (Blood 2005 Abramson, J.)
include cyclophosphamide, doxorubicin, vincristine, prednisone,
anti-CD20 monoclonal antibodies, etoposide, bleomycin, many of the
agents listed for Waldenstrom's, and any combination thereof, such
as ICE and R-ICE.
Examples of other therapeutic agents used to treat chronic
lymphocytic leukemia (CLL) (Spectrum, 2006, Fernandes, D.) include
idelalisib (Zydelig.RTM.), Chlorambucil (Leukeran),
Cyclophosphamide (Cyloxan, Endoxan, Endoxana, Cyclostin),
Fludarabine (Fludara), Pentstatin (Nipent), Cladribine (Leustarin),
Doxorubicin (Adriamycin.RTM., Adriblastine), Vincristine (Oncovin),
Prednisone, Prednisolone, Alemtuzumab (Campath, MabCampath), many
of the agents listed for Waldenstrom's, and combination
chemotherapy and chemoimmunotherapy, including the common
combination regimen: CVP (cyclophosphamide, vincristine,
prednisone); R-CVP (rituximab-CVP); ICE (iphosphamide, carboplatin,
etoposide); R-ICE (rituximab-ICE); FCR (fludarabine,
cyclophosphamide, rituximab); and FR (fludarabine, rituximab).
In another aspect, provided is a method of sensitizing a subject
(e.g., a human) who is (i) refractory to at least one chemotherapy
treatment, or (ii) in relapse after treatment with chemotherapy, or
both (i) and (ii), wherein the method comprises administering a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, or a pharmaceutical composition thereof, to the
subject. A subject who is sensitized is a subject who is responsive
to the treatment involving administration of the compound of
Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, or who has not developed resistance to such treatment.
In another aspect, provided herein are methods for treating a
subject (e.g., a human) for a cancer, with comorbidity, wherein the
treatment is also effective in treating the comorbidity. A
"comorbidity" to cancer is a disease that occurs at the same time
as the cancer.
In some embodiments, provided herein are methods for treating a
subject (e.g., a human) for chronic lymphocytic leukemia (CLL),
with comorbidity, wherein the treatment is also effective in
treating the comorbidity. Many subjects with CLL will have one or
more other diseases, for example diseases affecting the blood
pressure system, vascular and heart systems, endocrine and
metabolic systems, genitourinary system, musculoskeletal system,
respiratory system, neurological system, upper and lower
gastrointestinal systems, psychiatric system, ear, nose and throat
systems, renal system, or liver system. Specific morbidities of CLL
include, but are not limited to, one or more other cancers (e.g.
breast, head and neck, lung, melanoma, non-Hodgkin's T-cell
lymphoma, prostate, colon, small intestine, gynecologic and urinary
tract), hypertension, hyperlipidemia, coronary artery disease,
peripheral vascular diseases, cardiomyopathy, vulvular heart
disease, atrial fibrillation, cerebrovascular disease (e.g.
transient ischemic attack, stroke), chronic obstructive pulmonary
disease, joint disease, peptic ulcer, inflammatory bowel disease,
psychiatric illness, thyroid disease, benign prostate hyperplasia,
diabetes mellitus, and osteoarthritis (Satram-Hoang et al., Journal
of Cancer Therapy, 2013; 4:1321-1329; Thurmes et al., Leukemia
& Lymphoma, 2008; 49(1):49-56).
In some embodiments, a method of treating a comorbidity of CLL in a
subject (e.g., a human), wherein the method comprises administering
a compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, or a pharmaceutical composition thereof, to the
subject. In some embodiments, the comorbidity is selected from the
group consisting of one or more other cancers (e.g. breast, head
and neck, lung, melanoma, non-Hodgkin's T-cell lymphoma, prostate,
colon, small intestine, gynecologic and urinary tract),
hypertension, hyperlipidemia, coronary artery disease, peripheral
vascular diseases, cardiomyopathy, vulvular heart disease, atrial
fibrillation, cerebrovascular disease (e.g. transient ischemic
attack, stroke), chronic obstructive pulmonary disease, joint
disease, peptic ulcer, inflammatory bowel disease, psychiatric
illness, thyroid disease, benign prostate hyperplasia, diabetes
mellitus, and osteoarthritis.
Monotherapy and Combination Therapies
Also provided are methods of treatment in which a compound of
Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, is the only active agent given to a subject and also
includes methods of treatment in which a compound of Formula I, or
a pharmaceutically acceptable salt or co-crystal thereof, is given
to a subject in combination with one or more additional active
agents. Both monotherapy and combination therapies are intended and
described for use in the methods detailed herein, such as in a
method of treating any of the diseases or conditions detailed
herein and for use with any subject detailed herein.
Monotherapy
In some embodiments, a method of treating cancer, an allergic
disorder and/or an autoimmune and/or inflammatory disease, and/or
an acute inflammatory reaction comprises administering to a subject
in need thereof an effective amount of a compound of Formula I, or
a pharmaceutically acceptable salt or co-crystal thereof, wherein
the subject is not undergoing therapy for the same disease or
condition with another agent or procedure.
In some embodiments where the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, is
administered as a monotherapy to the subject who has been diagnosed
with or is suspected of having a cancer, the subject may be a human
who is (i) refractory to at least one anti-cancer therapy, or (ii)
in relapse after treatment with at least one anti-cancer therapy,
or both (i) and (ii). In some of embodiments, the subject is
refractory to at least two, at least three, or at least four
anti-cancer therapies (including, for example, standard or
experimental chemotherapies). For example, in some embodiments, the
subject may be a human who is (i) refractory to a therapy using an
anti-CD20 antibody, an alkylating agent (e.g., bendamustine), a
purine analog (e.g., fludarabine), an anthracycline, or any
combination thereof; (ii) in relapse after treatment with an
anti-CD20 antibody, an alkylating agent (e.g., bendamustine), a
purine analog (e.g., fludarabine), an anthracycline, or any
combination thereof, or both (i) and (ii).
A human subject who is refractory to at least one anti-cancer
therapy and/or is in relapse after treatment with at least one
anti-cancer therapy, as described above, may have undergone one or
more prior therapies. In some embodiments, such subjects have
undergone one, two, three, or four, or at least one, at least two,
at least three, at least four, or at least five, or between one and
ten, between one and nine, between one and eight, between one and
seven, between one and six, between one and five, or between one
and four, anti-cancer therapies prior to treatment using the
methods described herein (e.g., prior to the administration of the
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, as a monotherapy).
It should be understood that when a subject (e.g. a human) is
treated with the compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, as a monotherapy, the
subject may also undergo one or more other therapies that are not
anti-cancer therapies.
In some embodiments, a method of treating a comorbidity of a
cancer, including but not limited to CLL, in a subject (e.g., a
human) who has been diagnosed with cancer, e.g. CLL, wherein the
method comprises administering a therapy to treat the comorbidity
in combination with a compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, or a pharmaceutical
composition thereof, to the subject. In some embodiments, the
comorbidity is selected from the group consisting of one or more
other cancers (e.g. breast, head and neck, lung, melanoma,
non-Hodgkin's T-cell lymphoma, prostate, colon, small intestine,
gynecologic and urinary tract), hypertension, hyperlipidemia,
coronary artery disease, peripheral vascular diseases,
cardiomyopathy, vulvular heart disease, atrial fibrillation,
cerebrovascular disease (e.g. transient ischemic attack, stroke),
chronic obstructive pulmonary disease, joint disease, peptic ulcer,
inflammatory bowel disease, psychiatric illness, thyroid disease,
benign prostate hyperplasia, diabetes mellitus, and
osteoarthritis.
Combination Therapies
In some embodiments, a method of treating cancer, an allergic
disorder and/or an autoimmune and/or inflammatory disease, and/or
an acute inflammatory reaction comprises administering to a subject
in need thereof an effective amount of a compound of Formula I, or
a pharmaceutically acceptable salt or co-crystal thereof, together
with a second active agent, which can be useful for treating a
cancer, an allergic disorder and/or an autoimmune and/or
inflammatory disease, and/or an acute inflammatory reaction. For
example the second agent may be an anti-inflammatory agent.
Treatment with the second active agent may be prior to, concomitant
with, or following treatment with a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof. In some
embodiments, a compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof is combined with another
active agent in a single dosage form. In one embodiment, the
invention provides a product comprising a compound of the Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof, and
an additional therapeutic agent as a combined preparation for
simultaneous, separate or sequential use in therapy, e.g. a method
of treating a cancer, an allergic disorder and/or an autoimmune
and/or inflammatory disease, and/or an acute inflammatory
reaction.
Provided herein are also methods of treatment in which the compound
of Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, administered to a subject (e.g., a human) who has been
diagnosed with or is suspected of having a cancer is given to the
subject in combination with one or more additional therapies,
including one or more of the anti-cancer therapies described above.
Thus, in some embodiments, the method for treating cancer in a
subject (e.g., a human) in need thereof, comprises administering to
the subject a therapeutically effective amount of a compound of
Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, or a pharmaceutical composition thereof, together with one
or more additional therapies, which can be useful for treating the
cancer. The one or more additional therapies may involve the
administration of one or more therapeutic agents. Suitable
anti-cancer therapeutics that may be used in combination with a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof include, but are not limited to, one or more
agents selected from the group consisting of chemotherapeutic
agents (e.g. mitomycin C, carboplatin, taxol, cisplatin,
paclitaxel, etoposide, doxorubicin), radiotherapeutic antitumor
agents, topoisomerase I inhibitors (e.g. camptothesin or
topotecan), topoisomerase II inhibitors (e.g. daunomycin and
etoposide), alkylating agents (e.g. cyclophosphamide, melphalan and
BCNU), tubulin directed agents (e.g. taxol and vinblastine), PI3K
inhibitors (e.g. compounds A, B, and C below), inhibitors of lysyl
oxidase-like 2, and biological agents (e.g. antibodies such as anti
CD20 antibody, IDEC 8, immunotoxins, and cytokines).
In some embodiments, the method for treating cancer in a subject
(e.g., a human) in need thereof, comprises administering to the
subject a therapeutically effective amount of a compound of Formula
I, or a pharmaceutically acceptable salt or co-crystal thereof, or
a pharmaceutical composition thereof with one or more additional
therapies selected from the group consisting of fludarabine,
rituximab, obinutuzumab, alemtuzumab, cyclophosphamide,
chlorambucil, doxorubicin, doxorubicin hydrochloride, vincristine,
vincristine sulfate, melphalan, busulfan, carmustine, prednisone,
prednisolone, dexamethasone, methotrexate, cytarabine,
mitoxantrone, mitoxantrone hydrochloride, bortezomib, temsirolimus,
carboplatin, etoposide, thalidomide, cisplatin, lumiliximab,
anti-TRAIL, bevacizumab, galiximab, epratuzumab, SGN-40, anti-CD74,
ofatumumab, ha20, PRO131921, CHIR-12.12, apolizumab, milatuzumab,
bevacizumab, yttrium-90-labeled ibritumomab tiuxetan, tositumomab,
iodine-131 tositumomab, iphosphamide, GTOP-99 vaccine, oblimersen,
Flavopiridol, PD0332991, R-roscovitine, Styryl sulphones,
Obatoclax, TRAIL, Anti-TRAIL DR4 and DR5 antibodies, Everolimus,
BMS-345541, Curcumin, Vorinostat, lenalidomide, Geldanamycin,
perifosine, sildenafil citrate, CC-5103, simvastatin, enzastaurin,
campath-1H, DT PACE, antineoplaston A10, antineoplaston AS2-1, beta
alethine, filgrastim, recombinant interferon alfa, dolastatin 10,
indium In 111 monoclonal antibody MN-14, anti-thymocyte globulin,
cyclosporine, mycophenolate mofetil, therapeutic allogeneic
lymphocytes, tacrolimus, thiotepa, paclitaxel, aldesleukin,
docetaxel, ifosfamide, mesna, recombinant interleukin-12,
recombinant interleukin-11, ABT-263, denileukin diftitox,
tanespimycin, everolimus, pegfilgrastim, vorinostat, alvocidib,
recombinant flt3 ligand, recombinant human thrombopoietin,
lymphokine-activated killer cells, amifostine trihydrate,
aminocamptothecin, irinotecan hydrochloride, caspofungin acetate,
clofarabine, epoetin alfa, nelarabine, pentostatin, sargramostim,
vinorelbine ditartrate, WT-1 analog peptide vaccine, WT1 126-134
peptide vaccine, fenretinide, ixabepilone, oxaliplatin, monoclonal
antibody CD19, monoclonal antibody CD20, omega-3 fatty acids,
octreotide acetate, motexafin gadolinium, arsenic trioxide,
tipifarnib, autologous human tumor-derived HSPPC-96, veltuzumab,
bryostatin 1, PEGylated liposomal hydrochloride, peripheral blood
stem cell transplantation, autologous hematopoietic stem cell
transplantation, autologous bone marrow transplantation, infusion
of stem cells, bone marrow ablation with stem cell support, in
vitro-treated peripheral blood stem cell transplantation, umbilical
cord blood transplantation, low-LET cobalt-60 gamma ray therapy,
bleomycin, conventional surgery, radiation therapy, and
nonmyeloablative allogeneic hematopoietic stem cell
transplantation.
In some embodiments, the one or more additional therapies involve
the use of a phosphatidylinositol 3-kinase (PI3K) inhibitor,
including for example, Compounds A, B or C, or a pharmaceutically
acceptable salt of such compounds. The structures of Compounds A, B
and C are provided below.
##STR00134##
In other embodiments of the methods described above involving the
use of the compound of Formula I, or a pharmaceutically acceptable
salt or co-crystal thereof, in combination with one or more
additional therapies, the one or more additional therapies is other
than a therapy using Compound A, Compound B, or Compound C, or a
pharmaceutically acceptable salt of such compounds. In one
embodiment of the methods described above involving the use of the
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, in combination with one or more additional
therapies, the one or more additional therapies is other than a
therapy using Compound A, or a pharmaceutically acceptable salt or
co-crystal thereof. In another embodiment of the methods described
above involving the use of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, in
combination with one or more additional therapies, the one or more
additional therapies is other than a therapy using Compound B, or a
pharmaceutically acceptable salt or co-crystal thereof. In yet
another embodiment of the methods described above involving the use
of the compound of Formula I, or a pharmaceutically acceptable salt
or co-crystal thereof, in combination with one or more additional
therapies, the one or more additional therapies is other than a
therapy using Compound C, or a pharmaceutically acceptable salt or
co-crystal thereof.
In other embodiments, the one or more additional therapeutic agent
may be an inhibitors of lysyl oxidase-like 2 (LOXL2) and a
substance that bind to LOXL2, including for example, a humanized
monoclonal antibody (mAb) with an immunoglobulin IgG4 isotype
directed against human LOXL2.
The compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, can be useful as chemosensitizing agents, and,
thus, can be useful in combination with other chemotherapeutic
drugs, in particular, drugs that induce apoptosis.
A method for increasing sensitivity of cancer cells to
chemotherapy, comprising administering to a subject (e.g., human)
undergoing chemotherapy a chemotherapeutic agent together with a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, or a pharmaceutical composition thereof, in an
amount sufficient to increase the sensitivity of cancer cells to
the chemotherapeutic agent is also provided herein. Examples of
other chemotherapeutic drugs that can be used in combination with
chemical entities described herein include topoisomerase I
inhibitors (camptothesin or topotecan), topoisomerase II inhibitors
(e.g. daunomycin and etoposide), alkylating agents (e.g.
cyclophosphamide, melphalan and BCNU), tubulin directed agents
(e.g. taxol and vinblastine), and biological agents (e.g.
antibodies such as anti CD20 antibody, IDEC 8, immunotoxins, and
cytokines). In one embodiment of the method for increasing
sensitivity of cancer cells to chemotherapy, the chemotherapeutic
agent is other than Compound A, or a pharmaceutically acceptable
salt or co-crystal thereof. In another embodiment of the method for
increasing sensitivity of cancer cells to chemotherapy, the
chemotherapeutic agent is other than Compound B, or a
pharmaceutically acceptable salt or co-crystal thereof. In yet
another embodiment of the method for increasing sensitivity of
cancer cells to chemotherapy, the chemotherapeutic agent is other
than Compound C, or a pharmaceutically acceptable salt or
co-crystal thereof.
In some embodiments, the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, or a
pharmaceutical composition thereof, is used in combination with
Rituxan.RTM. (Rituximab) or other agents that work by selectively
depleting CD20+ B-cells.
Included herein are methods of treating cancer, an allergic
disorder and/or an autoimmune and/or inflammatory disease, and/or
an acute inflammatory reaction comprising administering to a
subject in need thereof an effective amount of a compound of
Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, or a pharmaceutical composition thereof, in combination
with an anti-inflammatory agent. Anti-inflammatory agents include
but are not limited to NSAIDs, non-specific and COX-2 specific
cyclooxgenase enzyme inhibitors, gold compounds, corticosteroids,
methotrexate, tumor necrosis factor receptor (TNF) receptors
antagonists, immunosuppressants and methotrexate. Examples of
NSAIDs include, but are not limited to ibuprofen, flurbiprofen,
naproxen and naproxen sodium, diclofenac, combinations of
diclofenac sodium and misoprostol, sulindac, oxaprozin, diflunisal,
piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen,
sodium nabumetone, sulfasalazine, tolmetin sodium, and
hydroxychloroquine. Examples of NSAIDs also include COX-2 specific
inhibitors (i.e., a compound that inhibits COX-2 with an IC50 that
is at least 50-fold lower than the IC50 for COX-1) such as
celecoxib, valdecoxib, lumiracoxib, etoricoxib and/or
rofecoxib.
In a further embodiment, the anti-inflammatory agent is a
salicylate. Salicylates include but are not limited to
acetylsalicylic acid or aspirin, sodium salicylate, and choline and
magnesium salicylates. The anti-inflammatory agent may also be a
corticosteroid. For example, the corticosteroid may be chosen from
cortisone, dexamethasone, methylprednisolone, prednisolone,
prednisolone sodium phosphate, and prednisone. In some embodiments,
the anti-inflammatory therapeutic agent is a gold compound such as
gold sodium thiomalate or auranofin. In some embodiments, the
anti-inflammatory agent is a metabolic inhibitor such as a
dihydrofolate reductase inhibitor, such as methotrexate or a
dihydroorotate dehydrogenase inhibitor, such as leflunomide.
In some embodiments, combinations in which at least one
anti-inflammatory compound is an anti-C5 monoclonal antibody (such
as eculizumab or pexelizumab), a TNF antagonist, such as
entanercept, or infliximab, which is an anti-TNF alpha monoclonal
antibody are used.
In some embodiments, combinations in which at least one therapeutic
agent is an immunosuppressant compound such as methotrexate,
leflunomide, cyclosporine, tacrolimus, azathioprine, or
mycophenolate mofetil are used.
Provided herein are also methods of treatment in which the compound
of Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, administered to a subject (e.g., a human) who has been
diagnosed with or is suspected of having an autoimmune disease is
given to the subject in combination with one or more
anti-inflammatory or immunosuppresant agents selected from the
group consisting of ibuprofen, flurbiprofen, naproxen, naproxen
sodium, diclofenac, diclofenac sodium, misoprostol, sulindac,
oxaprozin, diflunisal, piroxicam, indomethacin, etodolac,
fenoprofen calcium, ketoprofen, sodium nabumetone, sulfasalazine,
tolmetin sodium, hydroxychloroquine, celecoxib, valdecoxib,
lumiracoxib, etoricoxib, rofecoxib, acetylsalicylic acid, sodium
salicylate, choline salicylate, magnesium salicylate, cortisone,
dexamethasone, methylprednisolone, prednisolone, prednisolone
sodium phosphate, prednisone, gold sodium thiomalate, auranofin,
methotrexate, dihydroorotate leflunomide, leflunomide,
cyclosporine, tacrolimus, azathioprine, mycophenolate mofetil,
eculizumab, pexelizumab, entanercept, and infliximab.
It should be understood that any combinations of the additional
therapeutic agents described above may be used, as if each and
every combination was individually listed. For example, in some
embodiments, the additional therapeutic agents include a PI3K
inhibitor and a LOXL2 inhibitor.
Pharmaceutical Compositions and Administration
Compounds of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, are usually administered in the form of
pharmaceutical compositions. This disclosure therefore provides
pharmaceutical compositions that contain, as the active ingredient,
one or more of the compounds described, or a pharmaceutically
acceptable salt, pharmaceutically acceptable co-crystal or
pharmaceutically acceptable ester thereof, and one or more
pharmaceutically acceptable vehicle, such as excipients, carriers,
including inert solid diluents and fillers, diluents, including
sterile aqueous solution and various organic solvents, permeation
enhancers, solubilizers and adjuvants. The pharmaceutical
compositions may be administered alone or in combination with other
therapeutic agents. Such compositions are prepared in a manner well
known in the pharmaceutical art (see, e.g., Remington's
Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa.
17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd
Ed. (G. S. Banker & C. T. Rhodes, Eds.)
The pharmaceutical compositions may be administered in either
single or multiple doses by any of the accepted modes of
administration of agents having similar utilities, for example as
described in those patents and patent applications incorporated by
reference, including rectal, buccal, intranasal and transdermal
routes, by intra-arterial injection, intravenously,
intraperitoneally, parenterally, intramuscularly, subcutaneously,
orally, topically, as an inhalant, or via an impregnated or coated
device such as a stent, for example, or an artery-inserted
cylindrical polymer.
One mode for administration is parenteral, particularly by
injection. The forms in which the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, may be
incorporated for administration by injection include aqueous or oil
suspensions, or emulsions, with sesame oil, corn oil, cottonseed
oil, or peanut oil, as well as elixirs, mannitol, dextrose, or a
sterile aqueous solution, and similar pharmaceutical vehicles.
Aqueous solutions in saline may also conventionally be used for
injection. Ethanol, glycerol, propylene glycol, liquid polyethylene
glycol, and the like (and suitable mixtures thereof), cyclodextrin
derivatives, and vegetable oils may also be employed. The proper
fluidity can be maintained, for example, by the use of a coating,
such as lecithin, by the maintenance of the required particle size
in the case of dispersion and by the use of surfactants. The
prevention of the action of microorganisms can be brought about by
various antibacterial and antifungal agents, for example, parabens,
chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
Sterile injectable solutions are prepared by incorporating a
compound according to the present disclosure in the required amount
in the appropriate solvent with various other ingredients as
enumerated above, as required, followed by filtered sterilization.
Generally, dispersions are prepared by incorporating the various
sterilized active ingredients into a sterile vehicle which contains
the basic dispersion medium and the required other ingredients from
those enumerated above. In the case of sterile powders for the
preparation of sterile injectable solutions, the preferred methods
of preparation are vacuum-drying and freeze-drying techniques which
yield a powder of the active ingredient plus any additional desired
ingredient from a previously sterile-filtered solution thereof. In
some embodiments, for parenteral administration, sterile injectable
solutions are prepared containing a therapeutically effective
amount, e.g., 0.1 to 1000 mg, of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof. It will be
understood, however, that the amount of the compound actually
administered usually will be determined by a physician, in the
light of the relevant circumstances, including the condition to be
treated, the chosen route of administration, the actual compound
administered and its relative activity, the age, weight, and
response of the individual subject, the severity of the subject's
symptoms, and the like.
Oral administration is another route for administration of the
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof. Administration may be via capsule or enteric
coated tablets, or the like. In making the pharmaceutical
compositions that include the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, the active
ingredient is usually diluted by an excipient and/or enclosed
within such a carrier that can be in the form of a capsule, sachet,
paper or other container. When the excipient serves as a diluent,
it can be in the form of a solid, semi-solid, or liquid material
(as above), which acts as a vehicle, carrier or medium for the
active ingredient. Thus, the compositions can be in the form of
tablets, pills, powders, lozenges, sachets, cachets, elixirs,
suspensions, emulsions, solutions, syrups, aerosols (as a solid or
in a liquid medium), ointments containing, for example, up to 10%
by weight of the active compound, soft and hard gelatin capsules,
sterile injectable solutions, and sterile packaged powders.
Some examples of suitable excipients in an oral formulation include
lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum
acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium
silicate, microcrystalline cellulose, polyvinylpyrrolidone,
cellulose, sterile water, syrup, and methyl cellulose. The
formulations can additionally include: lubricating agents such as
talc, magnesium stearate, and mineral oil; wetting agents;
emulsifying and suspending agents; preserving agents such as methyl
and propylhydroxy-benzoates; sweetening agents; and flavoring
agents.
The pharmaceutical compositions as described herein can be
formulated so as to provide quick, sustained or delayed release of
the active ingredient after administration to the subject by
employing procedures known in the art. Controlled release drug
delivery systems for oral administration include osmotic pump
systems and dissolutional systems containing polymer-coated
reservoirs or drug-polymer matrix formulations. Examples of
controlled release systems are given in U.S. Pat. Nos. 3,845,770;
4,326,525; 4,902,514; and 5,616,345. Another formulation for use in
the methods of the present disclosure employs transdermal delivery
devices (patches). Such transdermal patches may be used to provide
continuous or discontinuous infusion of the compounds of the
present disclosure in controlled amounts. The construction and use
of transdermal patches for the delivery of pharmaceutical agents is
well known in the art. See, e.g., U.S. Pat. Nos. 5,023,252,
4,992,445 and 5,001,139. Such patches may be constructed for
continuous, pulsatile, or on demand delivery of pharmaceutical
agents.
In some embodiments, the compositions described herein are
formulated in a unit dosage form. The term "unit dosage forms"
refers to physically discrete units suitable as unitary dosages for
human subjects and other mammals, each unit containing a
predetermined quantity of active material calculated to produce the
desired therapeutic effect, in association with a suitable
pharmaceutical excipient (e.g., a tablet, capsule, ampoule). The
compounds are generally administered in a pharmaceutically
effective amount. In some embodiments, for oral administration,
each dosage unit contains from about 1 mg to about 5000 mg, about 1
mg to about 4000 mg, about 1 mg to about 3000 mg, about 1 mg to
about 2000 mg, about 2 mg to about 2000 mg, about 5 mg to about
2000 mg, about 10 mg to about 2000 mg, about 1 mg to about 1000 mg,
about 2 mg to about 1000 mg, about 5 mg to about 1000 mg, about 10
mg to about 1000 mg, about 25 mg to about 1000 mg, about 50 mg to
about 1000 mg, about 75 mg to about 1000 mg, about 100 mg to about
1000 mg, about 125 mg to about 1000 mg, about 150 mg to about 1000
mg, about 175 mg to about 1000 mg, about 200 mg to about 1000 mg,
about 225 mg to about 1000 mg, about 250 mg to about 1000 mg, about
300 mg to about 1000 mg, about 350 mg to about 1000 mg, about 400
mg to about 1000 mg, about 450 mg to about 1000 mg, about 500 mg to
about 1000 mg, about 550 mg to about 1000 mg, about 600 mg to about
1000 mg, about 650 mg to about 1000 mg, about 700 mg to about 1000
mg, about 750 mg to about 1000 mg, about 800 mg to about 1000 mg,
about 850 mg to about 1000 mg, about 900 mg to about 1000 mg, about
950 mg to about 1000 mg, about 1 mg to about 750 mg, about 2 mg to
about 750 mg, about 5 mg to about 750 mg, about 10 mg to about 750
mg, about 25 mg to about 750 mg, about 50 mg to about 750 mg, about
75 mg to about 750 mg, about 100 mg to about 750 mg, about 125 mg
to about 750 mg, about 150 mg to about 750 mg, about 175 mg to
about 750 mg, about 200 mg to about 750 mg, about 225 mg to about
750 mg, about 250 mg to about 750 mg, about 300 mg to about 750 mg,
about 350 mg to about 750 mg, about 400 mg to about 750 mg, about
450 mg to about 750 mg, about 500 mg to about 750 mg, about 550 mg
to about 750 mg, about 600 mg to about 750 mg, about 650 mg to
about 750 mg, about 700 mg to about 750 mg, about 1 mg to about 500
mg, about 2 mg to about 500 mg, about 5 mg to about 500 mg, about
10 mg to about 500 mg, about 25 mg to about 500 mg, about 50 mg to
about 500 mg, about 75 mg to about 500 mg, about 100 mg to about
500 mg, about 125 mg to about 500 mg, about 150 mg to about 500 mg,
about 175 mg to about 500 mg, about 200 mg to about 500 mg, about
225 mg to about 500 mg, about 250 mg to about 500 mg, about 300 mg
to about 500 mg, about 350 mg to about 500 mg, about 400 mg to
about 500 mg, about 450 mg to about 500 mg, about 1 mg to about 400
mg, about 2 mg to about 400 mg, about 5 mg to about 400 mg, about
10 mg to about 400 mg, about 25 mg to about 400 mg, about 50 mg to
about 400 mg, about 75 mg to about 400 mg, about 100 mg to about
400 mg, about 125 mg to about 400 mg, about 150 mg to about 400 mg,
about 175 mg to about 400 mg, about 200 mg to about 400 mg, about
225 mg to about 400 mg, about 250 mg to about 400 mg, about 300 mg
to about 400 mg, about 350 mg to about 400 mg, about 1 mg to about
300 mg, about 2 mg to about 300 mg, about 5 mg to about 300 mg,
about 10 mg to about 300 mg, about 25 mg to about 300 mg, about 50
mg to about 300 mg, about 75 mg to about 300 mg, about 100 mg to
about 300 mg, about 125 mg to about 300 mg, about 150 mg to about
300 mg, about 175 mg to about 300 mg, about 200 mg to about 300 mg,
about 225 mg to about 300 mg, about 250 mg to about 300 mg, about 1
mg to about 250 mg, about 2 mg to about 250 mg, about 5 mg to about
250 mg, about 10 mg to about 250 mg, about 25 mg to about 250 mg,
about 50 mg to about 250 mg, about 75 mg to about 250 mg, about 100
mg to about 250 mg, about 125 mg to about 250 mg, about 150 mg to
about 250 mg, about 175 mg to about 250 mg, about 200 mg to about
250 mg, about 225 mg to about 250 mg, about 1 mg to about 225 mg,
about 2 mg to about 225 mg, about 5 mg to about 225 mg, about 10 mg
to about 225 mg, about 25 mg to about 225 mg, about 50 mg to about
225 mg, about 75 mg to about 225 mg, about 100 mg to about 225 mg,
about 125 mg to about 225 mg, about 150 mg to about 225 mg, about
175 mg to about 225 mg, about 200 mg to about 225 mg, about 1 mg to
about 200 mg, about 2 mg to about 200 mg, about 5 mg to about 200
mg, about 10 mg to about 200 mg, about 25 mg to about 200 mg, about
50 mg to about 200 mg, about 75 mg to about 200 mg, about 100 mg to
about 200 mg, about 125 mg to about 200 mg, about 150 mg to about
200 mg, about 175 mg to about 200 mg, about 1 mg to about 175 mg,
about 2 mg to about 175 mg, about 5 mg to about 175 mg, about 10 mg
to about 175 mg, about 25 mg to about 175 mg, about 50 mg to about
175 mg, about 75 mg to about 175 mg, about 100 mg to about 175 mg,
about 125 mg to about 175 mg, about 150 mg to about 175 mg, about 1
mg to about 150 mg, about 2 mg to about 150 mg, about 5 mg to about
150 mg, about 10 mg to about 150 mg, about 25 mg to about 150 mg,
about 50 mg to about 150 mg, about 75 mg to about 150 mg, about 100
mg to about 150 mg, about 125 mg to about 150 mg, about 1 mg to
about 125 mg, about 2 mg to about 125 mg, about 5 mg to about 125
mg, about 10 mg to about 125 mg, about 25 mg to about 125 mg, about
50 mg to about 125 mg, about 75 mg to about 125 mg, about 100 mg to
about 125 mg, about 1 mg to about 100 mg, about 2 mg to about 100
mg, about 5 mg to about 100 mg, about 10 mg to about 100 mg, about
25 mg to about 100 mg, about 50 mg to about 100 mg, or about 75 mg
to about 100 mg of a compound of Formula I, about or a
pharmaceutically acceptable salt or co-crystal thereof.
In some embodiments, for oral administration, each dosage unit
contains about 1 mg, about 2 mg, about 5 mg, about 10 mg, about 15
mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40
mg, about 45 mg, about 50 mg, about 75 mg, about 100 mg, about 125
mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about
250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg,
about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700
mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about
950 mg, or about 1000 mg of a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof.
The dosages for oral administration described above may be
administered once daily (QD) or twice daily (BID). In some
embodiments, the compound of formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, or a pharmaceutical
composition thereof, is administered orally at a unit dosage of
about 1 mg QD, about 2 mg QD, about 5 mg QD, about 10 mg QD, about
15 mg QD, about 20 mg QD, about 25 mg QD, about 30 mg QD, about 35
mg QD, about 40 mg QD, about 45 mg QD, about 50 mg QD, about 75 mg
QD, about 100 mg QD, about 125 mg QD, about 150 mg QD, about 175 mg
QD, about 200 mg QD, about 225 mg QD, about 250 mg QD, about 300 mg
QD, about 350 mg QD, about 400 mg QD, about 450 mg QD, about 500 mg
QD, about 550 mg QD, about 600 mg QD, about 650 mg QD, about 700 mg
QD, about 750 mg QD, about 800 mg QD, about 850 mg QD, about 900 mg
QD, about 950 mg QD, or about 1000 mg QD. In some embodiments, the
compound of formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, or a pharmaceutical composition thereof, is
administered orally at a unit dosage of about 1 mg BID, about 2 mg
BID, about 5 mg BID, about 10 mg BID, about 15 mg BID, about 20 mg
BID, about 25 mg BID, about 30 mg BID, about 35 mg BID, about 40 mg
BID, about 45 mg BID, about 50 mg BID, about 75 mg BID, about 100
mg BID, about 125 mg BID, about 150 mg BID, about 175 mg BID, about
200 mg BID, about 225 mg BID, about 250 mg BID, about 300 mg BID,
about 350 mg BID, about 400 mg BID, about 450 mg BID, about 500 mg
BID, about 550 mg BID, about 600 mg BID, about 650 mg BID, about
700 mg BID, about 750 mg BID, about 800 mg BID, about 850 mg BID,
about 900 mg BID, about 950 mg BID, or about 1000 mg BID.
In some embodiments, for parenteral administration, each dosage
unit contains from 0.1 mg to 1 g, 0.1 mg to 700 mg, or 0.1 mg to
100 mg of a compound of Formula I, or a pharmaceutically acceptable
salt or co-crystal thereof.
For any of the dosage units as described herein, it will be
understood, however, that the amount of the compound actually
administered usually will be determined by a physician, in the
light of the relevant circumstances, including the condition to be
treated, the chosen route of administration, the actual compound
administered and its relative activity, the age, weight, and
response of the individual subject, the severity of the subject's
symptoms, and the like.
For preparing solid compositions such as tablets, the principal
active ingredient is mixed with a pharmaceutical excipient to form
a solid preformulation composition containing a homogeneous mixture
of the compound of Formula I, or a pharmaceutically acceptable salt
or co-crystal thereof. When referring to these preformulation
compositions as homogeneous, it is meant that the active ingredient
is dispersed evenly throughout the composition so that the
composition may be readily subdivided into equally effective unit
dosage forms such as tablets, pills and capsules.
The tablets or pills as described herein may be coated or otherwise
compounded to provide a dosage form affording the advantage of
prolonged action, or to protect from the acid conditions of the
stomach. For example, the tablet or pill can comprise an inner
dosage and an outer dosage component, the latter being in the form
of an envelope over the former. The two components can be separated
by an enteric layer that serves to resist disintegration in the
stomach and permit the inner component to pass intact into the
duodenum or to be delayed in release. A variety of materials can be
used for such enteric layers or coatings, such materials including
a number of polymeric acids and mixtures of polymeric acids with
such materials as shellac, cetyl alcohol, and cellulose
acetate.
Compositions for inhalation or insufflation may include solutions
and suspensions in pharmaceutically acceptable, aqueous or organic
solvents, or mixtures thereof, and powders. The liquid or solid
compositions comprising the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, may contain
suitable pharmaceutically acceptable excipients as described supra.
Preferably, the compositions are administered by the oral or nasal
respiratory route for local or systemic effect. Compositions in
preferably pharmaceutically acceptable solvents may be nebulized by
use of inert gases. Nebulized solutions may be inhaled directly
from the nebulizing device or the nebulizing device may be attached
to a facemask tent, or intermittent positive pressure breathing
machine. Solution, suspension, or powder compositions may be
administered, preferably orally or nasally, from devices that
deliver the formulation in an appropriate manner.
Dosing Regimen
In the methods provided herein, the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, or a
pharmaceutical composition thereof, is administered in a
therapeutically effective amount to achieve its intended purpose.
Determination of a therapeutically effective amount is well within
the capability of those skilled in the art, especially in light of
the detailed disclosure provided herein. In some embodiments
(methods of treating cancer), a therapeutically effective amount of
the compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, may (i) reduce the number of cancer cells; (ii)
reduce tumor size; (iii) inhibit, retard, slow to some extent, and
preferably stop cancer cell infiltration into peripheral organs;
(iv) inhibit (e.g., slow to some extent and preferably stop) tumor
metastasis; (v) inhibit tumor growth; (vi) delay occurrence and/or
recurrence of a tumor; and/or (vii) relieve to some extent one or
more of the symptoms associated with the cancer. In various
embodiments, the amount is sufficient to ameliorate, palliate,
lessen, and/or delay one or more of symptoms of cancer.
The therapeutically effective amount may vary depending on the
subject, and disease or condition being treated, the weight and age
of the subject, the severity of the disease or condition, and the
manner of administering, which can readily be determined by one or
ordinary skill in the art.
The dosing regimen of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, in the
methods provided herein may vary depending upon the indication,
route of administration, and severity of the condition, for
example. Depending on the route of administration, a suitable dose
can be calculated according to body weight, body surface area, or
organ size. The final dosing regimen is determined by the attending
physician in view of good medical practice, considering various
factors that modify the action of drugs, e.g., the specific
activity of the compound, the identity and severity of the disease
state, the responsiveness of the subject, the age, condition, body
weight, sex, and diet of the subject, and the severity of any
infection. Additional factors that can be taken into account
include time and frequency of administration, drug combinations,
reaction sensitivities, and tolerance/response to therapy. Further
refinement of the doses appropriate for treatment involving any of
the formulations mentioned herein is done routinely by the skilled
practitioner without undue experimentation, especially in light of
the dosing information and assays disclosed, as well as the
pharmacokinetic data observed in human clinical trials. Appropriate
doses can be ascertained through use of established assays for
determining concentration of the agent in a body fluid or other
sample together with dose response data.
The formulation and route of administration chosen may be tailored
to the individual subject, the nature of the condition to be
treated in the subject, and generally, the judgment of the
attending practitioner. For example, the therapeutic index of the
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, may be enhanced by modifying or derivatizing
the compound for targeted delivery to cancer cells expressing a
marker that identifies the cells as such. For example, the
compounds can be linked to an antibody that recognizes a marker
that is selective or specific for cancer cells, so that the
compounds are brought into the vicinity of the cells to exert their
effects locally, as previously described. See e.g., Pietersz et
al., Immunol. Rev., 129:57 (1992); Trail et al., Science, 261:212
(1993); and Rowlinson-Busza et al., Curr. Opin. Oncol., 4:1142
(1992).
The therapeutically effective amount of the compound of Formula I,
or a pharmaceutically acceptable salt or co-crystal thereof, may be
provided in a single dose or multiple doses to achieve the desired
treatment endpoint. As used herein, "dose" refers to the total
amount of an active ingredient (e.g., the compound of Formula I, or
a pharmaceutically acceptable salt or co-crystal thereof) to be
taken each time by a subject (e.g., a human). The dose
administered, for example for oral administration described above,
may be administered once daily (QD), twice daily (BID), three times
daily, four times daily, or more than four times daily. In some
embodiments, the dose of a compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, is
administered once daily. In some embodiments, the dose of a
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, is administered twice daily.
In some embodiments, exemplary doses of the compound of Formula I,
or a pharmaceutically acceptable salt or co-crystal thereof, for a
human subject may be from about 1 mg to about 5000 mg, about 1 mg
to about 4000 mg, about 1 mg to about 3000 mg, about 1 mg to about
2000 mg, about 2 mg to about 2000 mg, about 5 mg to about 2000 mg,
about 10 mg to about 2000 mg, about 1 mg to about 1000 mg, about 2
mg to about 1000 mg, about 5 mg to about 1000 mg, about 10 mg to
about 1000 mg, about 25 mg to about 1000 mg, about 50 mg to about
1000 mg, about 75 mg to about 1000 mg, about 100 mg to about 1000
mg, about 125 mg to about 1000 mg, about 150 mg to about 1000 mg,
about 175 mg to about 1000 mg, about 200 mg to about 1000 mg, about
225 mg to about 1000 mg, about 250 mg to about 1000 mg, about 300
mg to about 1000 mg, about 350 mg to about 1000 mg, about 400 mg to
about 1000 mg, about 450 mg to about 1000 mg, about 500 mg to about
1000 mg, about 550 mg to about 1000 mg, about 600 mg to about 1000
mg, about 650 mg to about 1000 mg, about 700 mg to about 1000 mg,
about 750 mg to about 1000 mg, about 800 mg to about 1000 mg, about
850 mg to about 1000 mg, about 900 mg to about 1000 mg, about 950
mg to about 1000 mg, about 1 mg to about 750 mg, about 2 mg to
about 750 mg, about 5 mg to about 750 mg, about 10 mg to about 750
mg, about 25 mg to about 750 mg, about 50 mg to about 750 mg, about
75 mg to about 750 mg, about 100 mg to about 750 mg, about 125 mg
to about 750 mg, about 150 mg to about 750 mg, about 175 mg to
about 750 mg, about 200 mg to about 750 mg, about 225 mg to about
750 mg, about 250 mg to about 750 mg, about 300 mg to about 750 mg,
about 350 mg to about 750 mg, about 400 mg to about 750 mg, about
450 mg to about 750 mg, about 500 mg to about 750 mg, about 550 mg
to about 750 mg, about 600 mg to about 750 mg, about 650 mg to
about 750 mg, about 700 mg to about 750 mg, about 1 mg to about 500
mg, about 2 mg to about 500 mg, about 5 mg to about 500 mg, about
10 mg to about 500 mg, about 25 mg to about 500 mg, about 50 mg to
about 500 mg, about 75 mg to about 500 mg, about 100 mg to about
500 mg, about 125 mg to about 500 mg, about 150 mg to about 500 mg,
about 175 mg to about 500 mg, about 200 mg to about 500 mg, about
225 mg to about 500 mg, about 250 mg to about 500 mg, about 300 mg
to about 500 mg, about 350 mg to about 500 mg, about 400 mg to
about 500 mg, about 450 mg to about 500 mg, about 1 mg to about 400
mg, about 2 mg to about 400 mg, about 5 mg to about 400 mg, about
10 mg to about 400 mg, about 25 mg to about 400 mg, about 50 mg to
about 400 mg, about 75 mg to about 400 mg, about 100 mg to about
400 mg, about 125 mg to about 400 mg, about 150 mg to about 400 mg,
about 175 mg to about 400 mg, about 200 mg to about 400 mg, about
225 mg to about 400 mg, about 250 mg to about 400 mg, about 300 mg
to about 400 mg, about 350 mg to about 400 mg, about 1 mg to about
300 mg, about 2 mg to about 300 mg, about 5 mg to about 300 mg,
about 10 mg to about 300 mg, about 25 mg to about 300 mg, about 50
mg to about 300 mg, about 75 mg to about 300 mg, about 100 mg to
about 300 mg, about 125 mg to about 300 mg, about 150 mg to about
300 mg, about 175 mg to about 300 mg, about 200 mg to about 300 mg,
about 225 mg to about 300 mg, about 250 mg to about 300 mg, about 1
mg to about 250 mg, about 2 mg to about 250 mg, about 5 mg to about
250 mg, about 10 mg to about 250 mg, about 25 mg to about 250 mg,
about 50 mg to about 250 mg, about 75 mg to about 250 mg, about 100
mg to about 250 mg, about 125 mg to about 250 mg, about 150 mg to
about 250 mg, about 175 mg to about 250 mg, about 200 mg to about
250 mg, about 225 mg to about 250 mg, about 1 mg to about 225 mg,
about 2 mg to about 225 mg, about 5 mg to about 225 mg, about 10 mg
to about 225 mg, about 25 mg to about 225 mg, about 50 mg to about
225 mg, about 75 mg to about 225 mg, about 100 mg to about 225 mg,
about 125 mg to about 225 mg, about 150 mg to about 225 mg, about
175 mg to about 225 mg, about 200 mg to about 225 mg, about 1 mg to
about 200 mg, about 2 mg to about 200 mg, about 5 mg to about 200
mg, about 10 mg to about 200 mg, about 25 mg to about 200 mg, about
50 mg to about 200 mg, about 75 mg to about 200 mg, about 100 mg to
about 200 mg, about 125 mg to about 200 mg, about 150 mg to about
200 mg, about 175 mg to about 200 mg, about 1 mg to about 175 mg,
about 2 mg to about 175 mg, about 5 mg to about 175 mg, about 10 mg
to about 175 mg, about 25 mg to about 175 mg, about 50 mg to about
175 mg, about 75 mg to about 175 mg, about 100 mg to about 175 mg,
about 125 mg to about 175 mg, about 150 mg to about 175 mg, about 1
mg to about 150 mg, about 2 mg to about 150 mg, about 5 mg to about
150 mg, about 10 mg to about 150 mg, about 25 mg to about 150 mg,
about 50 mg to about 150 mg, about 75 mg to about 150 mg, about 100
mg to about 150 mg, about 125 mg to about 150 mg, about 1 mg to
about 125 mg, about 2 mg to about 125 mg, about 5 mg to about 125
mg, about 10 mg to about 125 mg, about 25 mg to about 125 mg, about
50 mg to about 125 mg, about 75 mg to about 125 mg, about 100 mg to
about 125 mg, about 1 mg to about 100 mg, about 2 mg to about 100
mg, about 5 mg to about 100 mg, about 10 mg to about 100 mg, about
25 mg to about 100 mg, about 50 mg to about 100 mg, or about 75 mg
to about 100 mg.
In some embodiments, exemplary doses of the compound of Formula I,
or a pharmaceutically acceptable salt or co-crystal thereof, for a
human subject may be about 1 mg, about 2 mg, about 5 mg, about 10
mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35
mg, about 40 mg, about 45 mg, about 50 mg, about 75 mg, about 100
mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about
225 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg,
about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650
mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about
900 mg, about 950 mg, about 1000 mg, about 1200 mg, about 1400 mg,
about 1600 mg, about 1800 mg, about 2000 mg, about 2200 mg, about
2400 mg, about 2600 mg, about 2800 mg, about 3000 mg, about 3200
mg, about 3400 mg, about 3600 mg, about 3800 mg, about 4000 mg,
about 4200 mg, about 4400 mg, about 4600 mg, about 4800 mg, or
about 5000 mg.
In other embodiments, the methods provided comprise continuing to
treat the subject (e.g., a human) by administering the doses of the
compound of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, at which clinical efficacy is achieved or
reducing the doses by increments to a level at which efficacy can
be maintained. In some embodiments, the methods provided comprise
administering to the subject (e.g., a human) an initial daily dose
of 100 mg to 1000 mg of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, and
administering subsequent daily doses of the compound of Formula I,
or a pharmaceutically acceptable salt or co-crystal thereof, over
at least 6 days, wherein each subsequent daily dose is increased by
50 mg to 400 mg. Thus, it should also be understood that the dose
of the compound of Formula I, or a pharmaceutically acceptable salt
or co-crystal thereof, may be increased by increments until
clinical efficacy is achieved. Increments of about 25 mg, about 50
mg, about 100 mg, or about 125 mg, or about 150 mg, or about 200
mg, or about 250 mg, or about 300 mg can be used to increase the
dose. The dose can be increased daily, every other day, two, three,
four, five or six times per week, or once per week.
The frequency of dosing will depend on the pharmacokinetic
parameters of the compound administered, the route of
administration, and the particular disease treated. The dose and
frequency of dosing may also depend on pharmacokinetic and
pharmacodynamic, as well as toxicity and therapeutic efficiency
data. For example, pharmacokinetic and pharmacodynamic information
about the compound of Formula I, or a pharmaceutically acceptable
salt or co-crystal thereof, can be collected through preclinical in
vitro and in vivo studies, later confirmed in humans during the
course of clinical trials. Thus, for the compound of Formula I, or
a pharmaceutically acceptable salt or co-crystal thereof, used in
the methods provided herein, a therapeutically effective dose can
be estimated initially from biochemical and/or cell-based assays.
Then, dosage can be formulated in animal models to achieve a
desirable circulating concentration range that modulates Syk
expression or activity. As human studies are conducted further
information will emerge regarding the appropriate dosage levels and
duration of treatment for various diseases and conditions.
Toxicity and therapeutic efficacy of the compound of Formula I, or
a pharmaceutically acceptable salt or co-crystal thereof, can be
determined by standard pharmaceutical procedures in cell cultures
or experimental animals, e.g., for determining the LD.sub.50 (the
dose lethal to 50% of the population) and the ED.sub.50 (the dose
therapeutically effective in 50% of the population). The dose ratio
between toxic and therapeutic effects is the "therapeutic index",
which typically is expressed as the ratio LD.sub.50/ED.sub.50.
Compounds that exhibit large therapeutic indices, i.e., the toxic
dose is substantially higher than the effective dose, are
preferred. The data obtained from such cell culture assays and
additional animal studies can be used in formulating a range of
dosage for human use. The doses of such compounds lies preferably
within a range of circulating concentrations that include the
ED.sub.50 with little or no toxicity.
The administration of the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, may be
administered under fed conditions. The term fed conditions or
variations thereof refers to the consumption or uptake of food, in
either solid or liquid forms, or calories, in any suitable form,
before or at the same time when the compounds or pharmaceutical
compositions thereof are administered. For example, the compound of
Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, may be administered to the subject (e.g., a human) within
minutes or hours of consuming calories (e.g., a meal). In some
embodiments, the compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, may be administered to the
subject (e.g., a human) within 5-10 minutes, about 30 minutes, or
about 60 minutes consuming calories.
Articles of Manufacture and Kits
Compositions (including, for example, formulations and unit
dosages) comprising the compound of Formula I, or a
pharmaceutically acceptable salt or co-crystal thereof, can be
prepared and placed in an appropriate container, and labeled for
treatment of an indicated condition. Accordingly, provided is also
an article of manufacture, such as a container comprising a unit
dosage form of the compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, and a label containing
instructions for use of the compounds. In some embodiments, the
article of manufacture is a container comprising a unit dosage form
of the compound of Formula I, or a pharmaceutically acceptable salt
or co-crystal thereof, and at least one pharmaceutically acceptable
vehicle. The article of manufacture may be a bottle, vial, ampoule,
single-use disposable applicator, or the like, containing the
pharmaceutical composition provided in the present disclosure. The
container may be formed from a variety of materials, such as glass
or plastic and in one aspect also contains a label on, or
associated with, the container which indicates directions for use
in the treatment of cancer or inflammatory conditions. It should be
understood that the active ingredient may be packaged in any
material capable of improving chemical and physical stability, such
as an aluminum foil bag. In some embodiments, diseases or
conditions indicated on the label can include, for example,
treatment of cancer.
Any pharmaceutical composition provided in the present disclosure
may be used in the articles of manufacture, the same as if each and
every composition were specifically and individually listed for use
in an article of manufacture.
Kits comprising a pharmaceutical composition comprising a compound
of Formula I, or a pharmaceutically acceptable salt or co-crystal
thereof, are also provided. For example, a kit can comprise unit
dosage forms of the compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, and a package insert
containing instructions for use of the composition in treatment of
a medical condition. In some embodiments, the kit comprises a unit
dosage form of the compound of Formula I, or a pharmaceutically
acceptable salt or co-crystal thereof, and at least one
pharmaceutically acceptable vehicle. The instructions for use in
the kit may be for treating a cancer, including, for example, a
hematologic malignancy. In some embodiments, the instructions are
directed to use of the pharmaceutical composition for the treatment
of cancer, such as leukemia or lymphoma, including relapsed and
refractory leukemia or lymphoma. In some embodiments, the
instructions for use in the kit may be for treating a hematologic
cancer selected from the group consisting of small lymphocytic
lymphoma, non-Hodgkin's lymphoma, indolent non-Hodgkin's lymphoma,
refractory iNHL, mantle cell lymphoma, follicular lymphoma,
lymphoplasmacytic lymphoma, marginal zone lymphoma, immunoblastic
large cell lymphoma, lymphoblastic lymphoma, Splenic marginal zone
B-cell lymphoma (+/-villous lymphocytes), Nodal marginal zone
lymphoma (+/-monocytoid B-cells), Extranodal marginal zone B-cell
lymphoma of mucosa-associated lymphoid tissue type, cutaneous
T-cell lymphoma, extranodal T-cell lymphoma, anaplastic large cell
lymphoma, angioimmunoblastic T-cell lymphoma, mycosis fungoides,
B-cell lymphoma, diffuse large B-cell lymphoma, Mediastinal large
B-cell lymphoma, Intravascular large B-cell lymphoma, Primary
effusion lymphoma, small non-cleaved cell lymphoma, Burkitt's
lymphoma, multiple myeloma, plasmacytoma, acute lymphocytic
leukemia, T-cell acute lymphoblastic leukemia, B-cell acute
lymphoblastic leukemia, B-cell prolymphocytic leukemia, acute
myeloid leukemia, chronic lymphocytic leukemia, juvenile
myelomonocytic leukemia, minimal residual disease, hairy cell
leukemia, primary myelofibrosis, secondary myelofibrosis, chronic
myeloid leukemia, myelodysplastic syndrome, myeloproliferative
disease, and Waldestrom's macroglobulinemia. In one embodiment, the
instructions for use in the kit may be for treating chronic
lymphocytic leukemia or non-Hodgkin's lymphoma. In one embodiment,
the NHL is diffuse large B-cell lymphoma, mantle cell lymphoma,
follicular lymphoma, small lymphocytic lymphoma, lymphoplasmacytic
lymphoma, and marginal zone lymphoma. In one embodiment, the
hematologic malignancy is indolent non-Hodgkin's lymphoma. In some
embodiments, diseases or conditions indicated on the label can
include, for example, treatment of cancer.
In some instances, the instructions are directed to use of the
pharmaceutical composition for the treatment of a solid tumor,
wherein the solid tumor is from a cancer selected from the group
consisting of pancreatic cancer, urological cancer, bladder cancer,
colorectal cancer, colon cancer, breast cancer, prostate cancer,
renal cancer, hepatocellular cancer, thyroid cancer, gall bladder
cancer, lung cancer (e.g. non-small cell lung cancer, small-cell
lung cancer), ovarian cancer, cervical cancer, gastric cancer,
endometrial cancer, esophageal cancer, head and neck cancer,
melanoma, neuroendocrine cancer, CNS cancer, brain tumors (e.g.,
glioma, anaplastic oligodendroglioma, adult glioblastoma
multiforme, and adult anaplastic astrocytoma), bone cancer, soft
tissue sarcoma, retinoblastomas, neuroblastomas, peritoneal
effusions, malignant pleural effusions, mesotheliomas, Wilms
tumors, trophoblastic neoplasms, hemangiopericytomas, Kaposi's
sarcomas, myxoid carcinoma, round cell carcinoma, squamous cell
carcinomas, esophageal squamous cell carcinomas, oral carcinomas,
cancers of the adrenal cortex, ACTH-producing tumors.
In some instances, the instructions are directed to use of the
pharmaceutical composition for the treatment of an allergic
disorder and/or an autoimmune and/or inflammatory disease, and/or
an acute inflammatory reaction. In some embodiments, the
instructions are directed to use of the pharmaceutical composition
for the treatment of an autoimmune disease. in some embodiments,
the instructions are directed to use of the pharmaceutical
composition for the treatment of an autoimmune disease selected
from the group consisting of systemic lupus erythematosus,
myestenia gravis, rheumatoid arthritis, acute disseminated
encephalomyelitis, idiopathic thrombocytopenic purpura, multiple
sclerosis, Sjoegren's syndrome, psoriasis, autoimmune hemolytic
anemia, asthma, ulcerative colitis, Crohn's disease, irritable
bowel disease, and chronic obstructive pulmonary disease. In some
embodiments, the autoimmune disease is selected from the group
consisting of asthma, rheumatoid arthritis, multiple sclerosis,
chronic obstructive pulmonary disease and systemic lupus
erythematosus.
Any pharmaceutical composition provided in the present disclosure
may be used in the kits, the same as if each and every composition
were specifically and individually listed for use a kit.
Synthesis
The compounds of the disclosure may be prepared using methods
disclosed herein and routine modifications thereof which will be
apparent given the disclosure herein and methods well known in the
art. Conventional and well-known synthetic methods may be used in
addition to the teachings herein. The synthesis of typical
compounds of Formula I, or a pharmaceutically acceptable salt or
co-crystal thereof, may be accomplished as described in the
following examples. If available, reagents may be purchased
commercially, e.g. from Sigma Aldrich or other chemical
suppliers.
General Syntheses
Typical embodiments of compounds in accordance with the present
disclosure may be synthesized using the general reaction schemes
described below. It will be apparent given the description herein
that the general schemes may be altered by substitution of the
starting materials with other materials having similar structures
to result in products that are correspondingly different.
Descriptions of syntheses follow to provide numerous examples of
how the starting materials may vary to provide corresponding
products. Given a desired product for which the substituent groups
are defined, the necessary starting materials generally may be
determined by inspection. Starting materials are typically obtained
from commercial sources or synthesized using published methods. For
synthesizing compounds which are embodiments of the present
disclosure, inspection of the structure of the compound to be
synthesized will provide the identity of each substituent group.
The identity of the final product will generally render apparent
the identity of the necessary starting materials by a simple
process of inspection, given the examples herein.
Synthetic Reaction Parameters
The compounds of this disclosure can be prepared from readily
available starting materials using, for example, the following
general methods and procedures. It will be appreciated that where
typical or preferred process conditions (i.e., reaction
temperatures, times, mole ratios of reactants, solvents, pressures,
etc.) are given, other process conditions can also be used unless
otherwise stated. Optimum reaction conditions may vary with the
particular reactants or solvent used, but such conditions can be
determined by one skilled in the art by routine optimization
procedures.
Additionally, as will be apparent to those skilled in the art,
conventional protecting groups may be necessary to prevent certain
functional groups from undergoing undesired reactions. Suitable
protecting groups for various functional groups as well as suitable
conditions for protecting and deprotecting particular functional
groups are well known in the art. For example, numerous protecting
groups are described in T. W. Greene and G. M. Wuts (1999)
Protecting Groups in Organic Synthesis, 3rd Edition, Wiley, New
York, and references cited therein.
Furthermore, the compounds of this disclosure may contain a chiral
center. Accordingly, if desired, such compounds can be prepared or
isolated as pure stereoisomers, i.e., as individual enantiomers or
as stereoisomer-enriched mixtures. All such stereoisomers (and
enriched mixtures) are included within the scope of this
disclosure, unless otherwise indicated. Pure stereoisomers (or
enriched mixtures) may be prepared using, for example, optically
active starting materials or stereoselective reagents well-known in
the art. Alternatively, racemic mixtures of such compounds can be
separated using, for example, chiral column chromatography, chiral
resolving agents, and the like.
The starting materials for the following reactions are generally
known compounds or can be prepared by known procedures or obvious
modifications thereof. For example, many of the starting materials
are available from commercial suppliers such as Aldrich Chemical
Co. (Milwaukee, Wis., USA). Others may be prepared by procedures or
obvious modifications thereof, described in standard reference
texts such as Fieser and Fieser's Reagents for Organic Synthesis,
Volumes 1-15 (John Wiley, and Sons, 1991), Rodd's Chemistry of
Carbon Compounds, Volumes 1-5, and Supplementals (Elsevier Science
Publishers, 1989) organic Reactions, Volumes 1-40 (John Wiley, and
Sons, 1991), March's Advanced Organic Chemistry, (John Wiley, and
Sons, 5.sup.th Edition, 2001), and Larock's Comprehensive Organic
Transformations (VCH Publishers Inc., 1989).
The terms "solvent," "inert organic solvent" or "inert solvent"
refer to a solvent inert under the conditions of the reaction being
described in conjunction therewith (including, for example,
benzene, toluene, acetonitrile, tetrahydrofuran ("THF"),
dimethylformamide ("DMF"), chloroform, methylene chloride (or
dichloromethane), diethyl ether, methanol, pyridine and the like).
Unless specified to the contrary, the solvents used in the
reactions of the present disclosure are inert organic solvents, and
the reactions are carried out under an inert gas, preferably
nitrogen.
The term "q.s." means adding a quantity sufficient to achieve a
stated function, e.g., to bring a solution to the desired volume
(i.e., 100%).
The following examples are included to demonstrate preferred
embodiments of the disclosure. It should be appreciated by those of
skill in the art that the techniques disclosed in the examples
which follow represent techniques discovered by the inventor to
function well in the practice of the disclosure, and thus can be
considered to constitute preferred modes for its practice. However,
those of skill in the art should, in light of the present
disclosure, appreciate that many changes can be made in the
specific embodiments which are disclosed and still obtain a like or
similar result without departing from the spirit and scope of the
disclosure.
List of Abbreviations and Acronyms
TABLE-US-00006 Abbreviation Meaning .degree. C. Degree Celsius anal
Analytical ATP Adenosine-5'-triphosphate ATX II Anemonia sulcata
toxin AcOH Acetic acid ACN Acetonitrile CAN Ceric ammonium nitrate
XDI 1,1'-carbonyldiimidazole CHO Chinese hamster ovary conc.
Concentrated d Doublet DABCO 1,4-Diazabicyclo[2.2.2]octane DAST
(Diethylamino)sulfur trifluoride dd Doublet of doublets DCE
1,2-dichloroethane DCM Dichloromethane DEAD Diethyl
azodicarboxylate DIPEA N,N-diisopropylethylamine DMAP
4-dimethylaminopyridine DME 1,2-dimethoxyethane DMF
Dimethylformamide DMSO Dimethyl sulfoxide dppf
1,1'-Bis(diphenylphosphino)ferrocene EA Ethyl alcohol ECF
Extracellular fluid EDTA Ethylenediaminetetraacetic acid EGTA
Ethylene glycol tetraacetic acid equiv/eq Equivalents ESI
Electrospray ionization Ac Acetate Et Ethyl EtOAc Ethyl Acetate g
Grams HEPES (4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid)
HATU 2-(7-Aza-1H-Benzotriazole -1-yl)-1,1,3,3- tetramethyluronium
hexafluorophosphate hERG human Ether-a-go-go Related Gene HMDS
hexamethyldisilazane(azide) HPLC High-performance liquid
chromatography h Hours Hz Hertz IPA Isopropyl alcohol IC.sub.50 The
half maximal inhibitory concentration IMR-32 Human neuroblastoma
cell line J Coupling constant Kg Kilogram kHz Kilohertz LAH Lithium
ammonium hydride LCMS/LC-MS Liquid chromatography-mass spectrometry
M Molar m multiplet m/z mass-to-charge ratio M+ Mass peak M + H
Mass peak plus hydrogen mCPBA 3-chloroperoxybenzoic acid Me Methyl
MeOH Methanol mg Milligram MHz Megahertz min/m Minute ml/mL
Milliliter mM Millimolar mmol Millimole nmol Nanomole mOsmol
Milliosmole MRM Magnetic Resonance Microscopy MS Mass spectroscopy
ms Millisecond mV Millivolt mw Microwave N Normal mol Mole NMP
N-methylpyrrolidinone NMR Nuclear magnetic resonance pA Picoamps Ph
Phenyl ppm Parts per million prep Preparative q.s. Quantity
sufficient to achieve a stated function Rf Retention factor RP
Reverse phase RT/rt Room temperature s Second s Singlet SEM
2-(Trimethylsilyl)ethoxymethyl t Triplet TB Tonic Block TEA
Triethylamine TFA Trifluoroacetic acid THF Tetrahydrofuran TLC Thin
layer chromatography TMS trimethylsilyl TTX Tetrodotoxin UDB Use
Dependent Block WT Wild type .delta. Chemical shift .mu.g Microgram
.mu.L/.mu.l Microliter .mu.M Micromolar .mu.m Micrometer .mu.mol
Micromole
EXAMPLES
Preparation of Common Intermediates
Intermediate 1.01. Preparation of tert-Butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phen-
yl)carbamate IV and tert-butyl
4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl(6-(tributylstannyl)imidazo[1,2-a]-
pyrazin-8-yl)carbamate V
##STR00135## ##STR00136##
1-(4-Nitrophenyl)-4-(oxetan-3-yl)piperazine I
In a 500 mL round bottom flask, 1-(oxetan-3-yl)piperazine (3.02 g,
21.26 mmoles), potassium carbonate (5.87 g, 42.52 mmoles),
1-fluoro-4-nitrobenzene (3.00 g, 21.26 mmoles) was combined in
acetonitrile (33 mL) and stirred under nitrogen overnight at
100.degree. C. The mixture was diluted with water (100 mL) and
extracted with DCM (100 mL.times.3), dried over anhydrous sodium
carbonate, filtered and the filtrate was concentrated. The residue
was dissolved in minimal DCM using a sonicator and crashed out with
hexane. The precipitate was filtered, washed with hexane and dried
to provide the title compound I.
4-(4-(Oxetan-3-yl)piperazin-1-yl)aniline II
In a hydrogenation vessel,
1-(4-nitrophenyl)-4-(oxetan-3-yl)piperazine I (4.70 g, 17.85
mmoles) was dissolved as much as possible in MeOH (26 mL) and DCM
(5 mL). Pd/C (10%) (2.85 g, 2.68 mmoles) was added and the reaction
was stored under nitrogen. The reaction was shaken on the Parr
hydrogenator at 45 PSI. After 15 minutes, the reaction was fully
recharged to 45 PSI and shaken for an additional hour. The material
was filtered over celite, washed with 25% MeOH/DCM and concentrated
to provide the title compound II.
6-Bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-
-amine III
To 4-(4-(oxetan-3-yl)piperazin-1-yl)aniline II (2.00 g, 8.57
mmoles), hunig's base (3.29 mL) and
6,8-dibromoimidazo[1,2-a]pyrazine (2.37 g, 8.57 mmoles) was added
in DMF (43 mL). The reaction was stirred at 85.degree. C. in a
pressure tube for overnight. The material was quenched with
saturated sodium bicarbonate, extracted with DCM (120 mL.times.3)
and the organic layers were combined and washed with water (120
mL.times.3), dried over anhydrous sodium carbonate and
concentrated. The crude material was purified using a 120 g Isco
column and eluted off using a stepwise gradient of 0-60% (10%
MeOH/DCM). The desired fractions were combined and concentrated to
provide the title compound III.
tert-Butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-
-1-yl)phenyl)carbamate IV
6-bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin--
8-amine III (1000 mg, 2.33 mmol), di-tert-butyl dicarbonate
(1016.72 mg, 4.66 mmol) and N,N-dimethylpyridin-4-amine (21.34 mg,
0.17 mmol) were stirred in DCM (1.01 ml) and refluxed at 65.degree.
C. for 3 h. The reaction was diluted with 100 mL of DCM, washed
with H2O (.times.3), dried, filtered and concentrated. The crude
material was dissolved in minimal DCM, loaded onto a preloaded
silica loader and eluted off a 40 g column using 0-30% MeOH/DCM
over 20 column volumes. The desired fractions were combined and
concentrated to provide the title compound. This compound is used
in Example 2.
tert-Butyl
4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl(6-(tributylstannyl)imid-
azo[1,2-a]pyrazin-8-yl)carbamate V
In a 350 mL p-tube, tert-butyl
6-bromoimidazo[1,2-a]pyrazin-8-yl(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl-
)carbamate IV (8150 mg, 15.39 mmol),
1,1,1,2,2,2-hexabutyldistannane (11.67 ml, 23.09 mmol),
tetrakis(triphenylphosphine)palladium (889.43 mg, 0.77 mmol), and
tetrabutylammonium iodide (5686.03 mg, 15.39 mmol) were combined in
dioxane (62 ml) and heated to 110.degree. C. overnight. According
to LCMS, no starting material remained. The reaction was absorbed
onto celite and eluted off a 160 g alumina column using a
0-10-20-30-100% (50% EtOAc/Hex-Hex) gradient holding at 50% for
10-15 column volumes over 50-60 column volumes to provide the title
compound V. This compound is used in Examples 1 and 2.
Intermediate 1.02. Preparation tert-butyl
(6-bromo-5-methylimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin--
1-yl)phenyl)carbamate X
##STR00137## ##STR00138##
6-Methylpyrazin-2-amine VI
To a solution of anhydrous zinc(II) chloride (26.3 g, 193 mmol) in
THF (150 mL) at 0.degree. C., was added 3M methyl magnesium bromide
in diethyl ether (129 mL) drop wise over a period of 1 h.
[1,3-bis(diphenylphosphino)propane]nickel(II) chloride (2.08 g,
3.85 mmol) was then added and the mixture allowed to warm to room
temperature. To the above mixture, a solution of
6-chloro-2-aminopyrazine (5.00 g, 38.6 mmol) in anhydrous THF (25
mL) was added and the reaction stirred, under a nitrogen
atmosphere, at reflux for 6 h. After this time, the mixture was
cooled to room temperature, then to 0.degree. C. and carefully
quenched with saturated aqueous ammonium chloride (50 mL). The
organic layer was separated and dried over sodium sulfate. The
drying agent was filtered and the filtrate concentrated under
reduced pressure to provide crude 6-methylpyrazin-2-amine VI, which
was used in the next step without purification: .sup.1H NMR (400
MHz, CDCl.sub.3) .delta.: 7.63 (s, 1H), 7.53 (s, 1H), 4.96 (bs,
2H), 2.16 (s, 3H).
3,5-Dibromo-6-methylpyrazin-2-amine VII
To a solution of 6-methylpyrazin-2-amine VI (2.00 g, 18.3 mmol) in
THF (40 mL) at 10.degree. C., was added N-bromosuccinimide (6.70 g,
37.6 mmol) portion wise over 15 min and the mixture allowed to warm
to room temperature while stirring. After 2 h, the reaction was
concentrated under reduced pressure and the resulting residue was
purified by column chromatography (silica, gradient, hexanes to
EtOAc) to provide 3,5-dibromo-6-methylpyrazin-2-amine VII: .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta.: 4.93 (bs, 2H), 2.38 (s, 3H).
6,8-Dibromo-5-methylimidazo[1,2-a]pyrazine VIII
A mixture of 2-bromo-1,1-diethoxyethane (3.21 mL, 20.7 mmol) and
48% aqueous hydrobromic acid (1.0 mL) was stirred at reflux for 2
h. The reaction was then cooled to room temperature and treated
with sodium bicarbonate until gas evolution ceased. The mixture was
filtered and the filtrate diluted with ethanol (15 mL). To this
mixture, 3,5-dibromo-6-methylpyrazin-2-amine VII (3.00 g, 11.2
mmol) was added and the reaction stirred at reflux for 16 h. After
this time, the reaction was cooled to room temperature and
concentrated under reduced pressure to a volume of approximately 10
mL. The suspension was filtered and the filter cake washed with
cold ethanol (5 mL). The filter cake was then taken into water (50
mL) and the pH was adjusted to .about.8 with potassium carbonate.
The resulting suspension was filtered and the filter cake dried to
a constant weight under vacuum to provide
6,8-dibromo-5-methylimidazo[1,2-a]pyrazine VIII: .sup.1H NMR (400
MHz, CDCl.sub.3) .delta.: 7.90 (s, 1H), 7.72 (s, 1H), 2.74 (s,
3H).
6-Bromo-5-methyl-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]-
pyrazin-8-amine IX
The compound IX was prepared from
6,8-dibromo-5-methylimidazo[1,2-a]pyrazine VIII using the method as
described for preparing
6-bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin--
8-amine III in Intermediate Example 1.01.
tert-Butyl
(6-bromo-5-methylimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)-
piperazin-1-yl)phenyl)carbamate X
The compound X was prepared from
6-bromo-5-methyl-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a-
]pyrazin-8-amine IX using the method as described for preparing
tert-butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phen-
yl)carbamate IV in Intermediate Example 1.01. This compound is used
in Example 4.
Synthesis of Examples 1-7
Example 1 Preparation of
6-(6-amino-5-methylpyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phen-
yl)imidazo[1,2-a]pyrazin-8-amine (1)
##STR00139## ##STR00140##
2-Bis(tert-butoxycarbonyl)amino-6-bromo-3-chloropyrazine XI
6-Bromo-3-chloropyrazin-2-amine (2000 mg, 9.59 mmol) was dissolved
in DCM (48 ml) followed by triethylamine (3.99 ml, 28.78 mmol),
di-tert-butyl dicarbonate (4188.12 mg, 19.19 mmol), and
N,N-dimethylpyridin-4-amine (87.91 mg, 0.72 mmol). The reaction was
allowed to stir at room temperature for overnight. The crude
material was washed with water, dried, filtered and concentrated.
The crude material was dissolved in minimal DCM and loaded onto a
25 g prepacked silica loader and eluted off a 40 g column using
0-30% MeOH/DCM. The title compound XI was isolated and identified
by LCMS and NMR. The product was a mix of mono and bis
boc-protected material, mainly bis boc-protected as observed by
NMR.
tert-Butyl
tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3--
yl)piperazin-1-yl)phenyl)amino)imidazo[1,2-a]pyrazin-6-yl)-3-chloropyrazin-
-2-yl)carbamate XII
tert-Butyl
4-(4-(Oxetan-3-yl)piperazin-1-yl)phenyl(6-(tributylstannyl)imidazo[1,2-a]-
pyrazin-8-yl)carbamate V (1000 mg, 1.4 mmol),
2-Bis(tert-butoxycarbonyl)amino-6-bromo-3-chloropyrazine XI (552
mg, 1.35 mmol), and PdCl.sub.2(PPh.sub.3).sub.2 (142.77 mg, 0.20
mmol), in 1,4-Dioxane (11.27 ml) was irradiated in the microwave
for 20 min at 140.degree. C. The reaction was absorbed onto celite
and eluted off a 40 g Gold Isco column using 0-10-100% (30%
MeOH/DCM) over 20 column volumes. Fractions 34-39 were collected
and concentrated. According to NMR, the title compound XII was
identified and isolated.
tert-Butyl
(6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)(4--
(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate XIII
In a microwave vial, tert-butyl
tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperaz-
in-1-yl)phenyl)amino)imidazo[1,2-a]pyrazin-6-yl)-3-chloropyrazin-2-yl)carb-
amate XII (300 mg, 0.44 mmol), methylboronic acid (794.39 mg, 13.27
mmol), tetrakis(triphenylphosphine)palladium (51.12 mg, 0.04 mmol),
and 2M Na.sub.2CO.sub.3 (0.44 ml) were combined in DME (1.77 ml)
and irradiated in the microwave for 20 min at 150.degree. C. The
reaction was worked up using 25% MeOH/DCM and water. The organic
layers were combined, dried, filtered and concentrated. The crude
material was loaded onto silica and eluted off a 40 g Gold column
using 0-5-15-25-50% (30% MeOH/DCM) over 45 column volumes. The
desired fractions were concentrated and provided tert-butyl
(6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-
-3-yl)piperazin-1-yl)phenyl)carbamate XIII as the minor product and
the desired final compound 1 as an inseparable mixture (208 mg
total) and were taken in to the TFA reaction.
6-(6-Amino-5-methylpyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)pheny-
l)imidazo[1,2-a]pyrazin-8-amine (1)
To a solution of tert-butyl
6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl(4-(4-(oxetan-3-
-yl)piperazin-1-yl)phenyl)carbamate XIII (48 mg, 0.09 mmol) and
6-(6-amino-5-methylpyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phen-
yl)imidazo[1,2-a]pyrazin-8-amine (1, 160 mg, 0.35 mmol) in DCM (2.5
ml) was added TFA (0.16 ml, 2.15 mmol). Additional TFA (0.48 ml,
6.5 mmol) was added to the reaction mixture to ensure reaction
completion. The reaction was then cooled to 0.degree. C. and
quenched with sat. NaHCO.sub.3, then extracted with DCM (5
ml.times.3), and the combined organic layers were washed with water
(5 ml.times.2), brine (5 ml.times.1), dried (Na.sub.2SO.sub.4), and
concentrated to give the crude product. The crude material was
absorbed onto silica and eluted off a 24 g Gold Isco column using
0-15-25-40-100% (30% MeOH/DCM). The desired fractions were combined
and concentrated to provide the desired compound. LCMS-ESI.sup.+
(m/z): [M+H].sup.+: 458.22. .sup.1H NMR (300 MHz, d.sub.6-DMSO)
.delta.: 9.48 (s, 1H), 8.54 (s, 1H), 8.41 (s, 1H), 8.11 (s, 1H),
7.95 (d, 2H), 7.6 (s, 1H), 6.98 (d, 2H), 6.2 (s, 2H), 4.58-4.45
(dt, 4H), 3.3 (m, 1H), 3.14 (t, 4H), 2.50-2.4 (dt, 4H), 2.33 (s,
1H). Alternatively, compound XII could be taken directly to this
step and similarly de-protected to provide the 5-chloropyrazine
substituted analog.
Example 2. Preparation of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine (2)
##STR00141##
2-Bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV
To a mixture of 6-bromopyrazin-2-amine (5 g, 28.7 mmol) and
di-tert-butyl dicarbonate (25.09 g, 114.94 mmol) was added DCM (10
ml) followed by DMAP (0.351 g, 29 mmol). The reaction was heated to
55.degree. C. for 1 h, cooled to RT, the reaction was partitioned
between water and DCM, purified on silica gel and concentrated to
provide of 2-bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+: 374.14. .sup.1H NMR (DMSO)
.delta.: 8.84 (d, 2H), 1.39 (s, 18H).
tert-Butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a-
]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate
XVI--CHEMISTRY A route
tert-Butyl
4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl(6-(tributylstannyl)imidazo[1,2-a]-
pyrazin-8-yl)carbamate V (215 mg, 0.291 mmol), was combined with
2-bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV (217.58 mg,
0.581 mmol), bis(triphenylphosphine)palladium(II) dichloride (30.61
mg, 0.044 mmol) and 1,4-dioxane (5 ml). The reaction mixture was
stirred in a microwave reactor at 120.degree. C. for 30 min. The
reaction mixture was quenched with saturated KF, extracted with
EtOAc, purified on silica gel, eluted with EtOAc. The desired
fractions were combined and concentrated to provide 100 mg (46%
yield) of tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate XVI.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+: 744.4. .sup.1H NMR (300 MHz
d.sub.6-DMSO) .delta.: 9.37 (s, 1H), 9.18 (s, 1H), 8.77 (s, 1H),
8.33 (d, 1H), 7.87 (d, 1H), 7.28-7.25 (d, 2H), 6.92-6.89 (d, 2H),
4.55-4.41 (m, 4H), 3.4 (m, 1H), 3.14-3.11 (m, 4H), 2.37-2.34 (m,
4H), 1.37 (s, 18H), 1.3 (s, 9H).
tert-Butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a-
]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate
XVI--CHEMISTRY B route
Step 1: To a dry 250 mL round-bottomed flask was added
2-bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV (1.0 g, 1.0
equiv, 2.67 mmol), KOAc (790 mg, 8.02 mmol, 3.0 equiv),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (750
mg, 2.94 mmol, 1.1 equiv), Pd(dba) (171 mg, 0.187 mmol, 0.07 equiv)
and X-phos (128 mg, 0.267 mmol, 0.1 equiv) followed by 1,4-dioxane
(25 mL) and the solution was sonicated for 5 min and then purged
with N.sub.2 gas for 5 min. The flask with contents was then placed
under N.sub.2 atmosphere and heated at 110.degree. C. for 90 min.
Once full conversion to the pinacolboronate was achieved by LCMS,
the reaction was removed from heat and allowed to cool to RT. Once
cool, the reaction contents were filtered through Celite and the
filter cake was washed 3.times.20 mL EtOAc. The resultant solution
was then concentrated down to a deep red-orange syrup providing N,
N-BisBoc
6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazin-2-amine XV,
which was used directly in the next step.
Step 2: The freshly formed N, N-BisBoc
6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazin-2-amine XV
(2.67 mmol based on 100% conversion, 2.0 equiv based on bromide)
was dissolved in 20 Ml of 1,2-dimethoxyethane and to that solution
was added tert-butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phen-
yl)carbamate IV (707 mg, 1.34 mmol, 1.0 equiv), Na.sub.2CO.sub.3
(283 mg, 2.67 mmol, 2.0 equiv), Pd(PPh.sub.3).sub.4 (155 mg, 0.134
mmol, 0.1 equiv) and water (10 mL) and the solution was degassed
for 5 min using N.sub.2 gas. The reaction was then placed under
N.sub.2 atmosphere and heated at 110.degree. C. for 90 min. LCMS
showed complete consumption of the bromide starting material and
the reaction was removed from heat and allowed to cool to RT. The
reaction was diluted with 100 mL water and 100 mL 20% MeOH/DCM and
the organic layer was recovered, extracted 1.times. sat.
NaHCO.sub.3, 1.times. sat brine and then dried over
Na.sub.2SO.sub.4. The solution was then filtered and concentrated
down to an orange-red solid. The sample was then slurried in warm
MeOH, sonicated then filtered, washing 2.times.20 mL with cold MeOH
and then the cream-colored solid was dried on hi-vacuum overnight
to yield 905 mg of tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate XVI.
6-(6-Aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo-
[1,2-a]pyrazin-8-amine (2)
To a solution of tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate XVI (200 mg,
0.269 mmol) in DCM (2 ml) was added TFA (0.5 ml, 6.578 mmol). The
reaction was stirred at rt for 16 h, saturated sodium bicarbonate
was added, extracted with EtOAC and purified on silica gel, eluted
with 5% MeOH/EtOAc, 20% MeOH/EtOAc. The desired fractions were
combined and concentrated to provide the title compound 2.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+: 444.2. .sup.1H NMR (300 MHz
d.sub.6-DMSO) .delta.: 9.5 (s, 1H), 8.588 (s, 1H), 8.47 (s, 1H),
8.12 (d, 1H), 7.95-7.92 (d, 2H), 7.88 (s, 1H), 7.62 (s, 1H),
6.99-6.96 (d, 2H), 6.46 (s, 2H), 4.57-4.53 (m, 2H), 4.48-4.44 (m,
2H), 3.43 (m, 1H), 3.15-3.12 (m, 4H), 2.41-2.38 (m, 4H).
Example 2--Alternate Synthesis
##STR00142##
Di-tert-butyl
{6-[8-({4-[4-(oxetan-3-yl)piperazin-1-yl]phenyl}amino)imidazo[11,2-a]pyra-
zin-6-yl]pyrazin-2-yl}imidodicarbonate
To a 720 L reactor, was added di-tert-butyl
(6-bromopyrazin-2-yl)imidodicarbonate (18.5 kg, 1.41 equiv, 49
mol), bis(pinacolato)diboron (13.8 kg, 1.56 equiv, 54 mol),
potassium propionate (11.9 kg, 3.02 equiv, 106 mol), and
bis(di-tert-butyl(4-dimethylaminophenyl)
phosphine)dichloropalladium (1.07 kg, 0.0043 equiv, 1.5 mol),
followed by degassed toluene (173 L). The mixture was degassed then
heated at 65.degree. C. until the reaction was deemed complete (0%
tert-butyl
2-((6-bromopyrazin-2-yl)(tert-butoxycarbonyl)amino)-2-oxoacetate)
by UPLC. Upon completion, the reaction was cooled to 23.degree. C.
Once cooled,
6-bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]-
pyrazin-8-amine (15.0 kg, 1.00 equiv, 35 mol) was added and the
mixture was degassed. A degassed aqueous potassium carbonate
solution prepared using water (54 L) and potassium carbonate (20.6
g, 4.26 equiv, 149 mol) was then added to the reaction mixture and
the reactor contents was degassed. The reactor contents was heated
at 65.degree. C. until reaction was deemed complete (1%
6-bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin--
8-amine) by UPLC. Upon completion, the reaction was cooled to
24.degree. C.
The cooled mixture was concentrated and then diluted with
dichloromethane (300 L), transferred to a 1900 L reactor and rinsed
forward with dichloromethane (57 L). N-acetyl-L-cysteine (3.8 kg)
was charged and the mixture was agitated for 15 h. Water (135 L)
was then added and the mixture was filtered and rinsed forward with
dichloromethane (68 L). The organic layer was recovered and washed
with a brine solution prepared using water (68 L) and sodium
chloride (7.5 kg).
The resultant organic layer was polish filtered then concentrated
and tert-butyl methyl ether (89.9 kg) was slowly charged keeping
the temperature at 31.degree. C. The contents was cooled to
0.degree. C. and aged, then filtered and rinsed with tert-butyl
methyl ether (32.7 kg) and dried at 40.degree. C. to give 17.2 kg
of di-tert-butyl
{6-[8-({4-[4-(oxetan-3-yl)piperazin-1-yl]phenyl}amino)imidazo[1,2-a]pyraz-
in-6-yl]pyrazin-2-yl}imidodicarbonate.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+: 644.3. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta.: 9.43 (s, 1H), 8.58 (s, 1H), 8.53 (s, 1H), 8.02
(s, 1H), 7.84 (m, 2H), 7.63 (d, 1H), 7.61 (d, 1H), 7.04 (m, 2H),
4.71 (m, 4H), 3.59 (m, 1H), 3.27 (m, 4H), 2.55 (m, 4H), 1.46 (s,
18H).
6-(6-Aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidazo-
[1,2-a]pyrazin-8-amine succinate (Example 2)
To a slurry of di-tert-butyl
{6-[8-({4-[4-(oxetan-3-yl)piperazin-1-yl]phenyl}amino)imidazo[1,2-a]pyraz-
in-6-yl]pyrazin-2-yl}imidodicarbonate (225 g, 0.35 mol, 1 mol eq.)
in water (12 parts) was added a solution of sulfuric acid (3.1
parts, 6.99 mol, 20 mol eq.) in water (5 parts). The reaction was
heated to ca. 40.degree. C. and stirred at this temperature for ca.
4 h at which point the reaction is deemed complete. The reaction
mixture was cooled to ca. 22.degree. C., acetone (3 parts) was
charged and a solution of sodium carbonate (4.1 parts, 8.75 mol,
25.0 mol eq.) in water (15 parts) was added. The resulting slurry
was filtered and the wet cake was washed with water in portions
(4.times.1 parts), then with tert-butyl methyl ether (4 parts). The
wet cake (Example 2 free base) was dried at ca. 60.degree. C. To
the slurry of dry Example 2 free base in 2-propanol (2.3 parts) was
added a solution of succinic acid (Based on the isolated Example 2
free base: 0.43 parts, 1.6 mol eq.) in 2-propanol (15 parts). The
resulting slurry was heated to ca. 40.degree. C. and stirred at
this temperature for ca. 2 h and then cooled to ca. 22.degree. C.,
followed by a stir period of ca. 16 h. The slurry was filtered at
ca. 22.degree. C. and the wet cake was washed with 2-propanol (5
parts) and dried at ca. 60.degree. C. to afford the product.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+: 620.65. .sup.1H NMR (400 MHz
d.sub.6-DMSO) .delta.:12.2 (broad s, 1.5H), 9.58 (s, 1H), 8.63 (s,
1H), 8.50 (s, 1H), 8.15 (s, 1H), 7.95 (d, 2H), 7.90 (s, 1H), 7.64
(s, 1H), 7.00 (d, 2H), 6.50 (s, 2H), 4.52 (dd, 4H), 3.45 (m, 1H),
3.19 (m, 4H), 2.40 (m, 10H).
Example 3. Preparation of
(R)-(4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)pheny-
l)morpholin-2-yl)methanol (3)
##STR00143## ##STR00144##
(R)-(4-(4-((6-Bromoimidazo[1,2-a]pyrazin-8-yl)amino)phenyl)morpholin-2-yl)-
methanol XVII
In a 250 mL round bottom flask equipped with a condenser was placed
6,8-dibromoimidazo[1,2-a]pyrazine (2000 mg, 7.22 mmol) and added 30
mL isopropanol followed by N,N-diisopropylethylamine (2.52 ml,
14.44 mmol) and (R)-(4-(4-aminophenyl)morpholin-2-yl)methanol
(1504.12 mg, 7.22 mmol). The reaction was heated to reflux (oil
bath 95.degree. C.) overnight. The reaction was cooled and
precipitates were collected by filtration and washed with
isopropanol followed by hexanes to give the desired compound
XVII.
(R)-tert-Butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(2-(((tert-butoxycarbonyl)oxy)methy-
l)morpholino)phenyl)carbamate XVIII
In a 250 mL round bottom flask was placed
(R)-(4-(4-((6-bromoimidazo[1,2-a]pyrazin-8-yl)amino)phenyl)morpholin-2-yl-
)methanol XVII (2.80 g, 6.9 mmol) and added DCM followed by
triethylamine (2.9 mL, 2.1 g, 20.8 mmol), DMAP (63 g, 0.52 mmol)
and di-tert-butyl dicarbonate (3.8 g, 17.3 mmol). The reaction was
stirred overnight then diluted with DCM and water, separated,
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The crude material was
purified by chromatography: ISCO 40 g silica with 25 g silica
loader, eluting with 0-100% EtOAc/hexanes to give compound
XVIII.
(R)-tert-Butyl
(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)(6-(tributylsta-
nnyl)imidazo[1,2-a]pyrazin-8-yl)carbamate XIX
(R)-tert-Butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(2-(((tert-butoxycarbonyl)oxy)methy-
l)morpholino)phenyl)carbamate XVIII was reacted according to the
analogous method of Example Intermediate 1.01 to provide
(R)-tert-butyl
(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)(6-(tributylsta-
nnyl)imidazo[1,2-a]pyrazin-8-yl)carbamate XIX.
(R)-tert-Butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
-yl)(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)carbamate
XX
(R)-tert-Butyl
(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)(6-(tributylsta-
nnyl)imidazo[1,2-a]pyrazin-8-yl)carbamate XIX was reacted with
2-Bis(tert-butoxycarbonyl)amino-6-bromopyrazine XIV according to
the analogous method of CHEMISTRY A as described in Example 2 to
provide the desired compound (R)-tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
-yl)(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)carbamate
XX.
(R)-(4-(4-((6-(6-Amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)ami-
no)phenyl)morpholin-2-yl)methanol (3)
(R)-tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
-yl)(4-(2-(((tert-butoxycarbonyl)oxy)methyl)morpholino)phenyl)carbamate
XX (460 mg, 0.56 mmol) in DCM was added to a round bottom flask,
and TFA (1.29 ml, 16.85 mmol) was added. The reaction was partially
complete after stirring .about.5 hours. Added an additional 10 eq
TFA and stirred overnight, then concentrated under reduced
pressure. 10% MeOH/DCM (.about.100 mL) and sat. aq. sodium
bicarbonate were added and stirred 15 min, separated, extracted
with .about.100 mL 10% MeOH/DCM. The organic layers were combined,
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure and dried under vacuum. The
resulting solid was triturated with DCM, collected solids via
filtration and dried under vacuum to give compound 3.
LCMS-ESI.sup.+ (m/z): [M+H].sup.+: 419.2. .sup.1H NMR (300 MHz
d.sub.6-DMSO) .delta.: 9.57 (s, 1H), 8.59 (s, 1H), 8.47 (s, 1H),
8.13 (d, J=1.2 Hz, 1H), 8.06-7.90 (m, 2H), 7.87 (s, 1H), 7.62 (d,
J=1.1 Hz, 1H), 7.05-6.93 (m, 2H), 6.49 (s, 2H), 4.78 (t, J=5.8 Hz,
1H), 3.98-3.87 (m, 1H), 3.71-3.36 (m, 7H), 2.63 (td, J=11.7, 3.4
Hz, 1H), 2.37 (dd, J=12.1, 10.5 Hz, 1H). The corresponding (S)
isomer, or racemic mixture of compounds is prepared similarly,
using (S)-(4-(4-aminophenyl)morpholin-2-yl)methanol or a racemic
mixture of (4-(4-aminophenyl)morpholin-2-yl)methanol, respectively,
in the first step.
Example 4. Preparation of
6-(6-aminopyrazin-2-yl)-5-methyl-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phen-
yl)imidazo[1,2-a]pyrazin-8-amine (4)
##STR00145##
tert-Butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)-5-methylimid-
azo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate
XXI
tert-Butyl
(6-bromo-5-methylimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin--
1-yl)phenyl)carbamate X was reacted with XV according to the
methods of CHEMISTRY B as described in Example 2 to provide the
desired compound XXI.
6-(6-aminopyrazin-2-yl)-5-methyl-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)pheny-
l)imidazo[1,2-a]pyrazin-8-amine (4)
The compound tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)-5-methylimidazo[1,2-a]-
pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)carbamate XXI
was de-protected by the analogous method described in Example 2 to
provide the desired compound 4. LCMS-ESI.sup.+ (m/z): [M+H].sup.+:
458.32. .sup.1H NMR (300 MHz, d.sub.6-DMSO) .delta.: 9.28 (s, 1H),
8.28 (s, 1H), 8.04 (s, 1H), 7.89 (d, 2H), 7.83 (s, 1H), 7.7 (s,
1H), 6.91 (d, 2H), 6.46 (s, 2H), 4.6-4.4 (dt, 4H), 3.43 (m, 1H),
3.1 (t, 4H), 2.49 (s, 3H), 2.4 (t, 4H).
Example 5. Preparation of
2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2-(4-(oxe-
tan-3-yl)piperazin-1-yl)phenoxy)ethanol (5)
##STR00146##
2-(2-(2-Fluoro-5-nitrophenoxy)ethoxy)tetrahydro-2H-pyran XXII
A mixture of 2-fluoro-5-nitrophenol (4 g, 25 mmol),
2-(2-bromoethoxy)tetrahydro-2H-pyran (4.4 mL, 28 mmol) and
potassium carbonate (4.2 g 30 mmol) in DMF (50 mL) was stirred at
50.degree. C. for 16 h. The reaction was cooled to room
temperature, diluted with EtOAc and H.sub.2O. The aqueous layer was
separated and extracted with EtOAc. The combined organic extracts
were washed with H.sub.2O (5.times.'s to remove DMF) and brine and
dried over sodium sulfate. The resulting residue was purified by
column chromatography ISCO Rf (40 g column) eluting with a gradient
of 100% hexanes--1:1 hexanes:EtOAc to provide
2-(2-(2-fluoro-5-nitrophenoxy)ethoxy)tetrahydro-2H-pyran XXII.
1-(4-Nitro-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)-4-(oxetan-3--
yl)piperazine XXIII
A mixture of
2-(2-(2-fluoro-5-nitrophenoxy)ethoxy)tetrahydro-2H-pyran XXII (1550
mg, 5.43 mmol), 1-(oxetan-3-yl)piperazine (772 mg, 5.43 mmol) and
potassium carbonate (1126.41 mg, 8.15 mmol) in NMP (6 mL) was
stirred at 100.degree. C. for 8 h. The aqueous layer was separated
and extracted with EtOAc. The combined organic extracts were washed
with H.sub.2O (5.times. to remove NMP) and brine and dried over
sodium sulfate. The resulting residue was purified by column
chromatography ISCO Rf (24 g column) eluting with a gradient of
100% DCM--60:35:5 DCM:Et.sub.2O:MeOH to provide
1-(4-nitro-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)-4-(oxetan-3-
-yl)piperazine XXIII.
4-(4-(Oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)etho-
xy)aniline XXIV
To a suspension of
1-(4-nitro-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)-4-(oxetan-3-
-yl)piperazine XXIII (2100 mg, 5.1 mmol) in ethanol (50 mL) was
added 10% Pd/C (50% wet, 390 mg dry weight) in a 500-mL Parr
hydrogenation bottle. The bottle was evacuated, charged with
hydrogen gas to a pressure of 50 psi and shaken at rt for 2 h on a
Parr hydrogenation apparatus. The reaction mixture was filtered,
and washed with ethanol. The filtrate was concentrated in vacuo to
give
4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)eth-
oxy)aniline XXIV.
6-Bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2
yl)oxy)ethoxy)phenyl)imidazo[1,2-a]pyrazin-8-amine XXV
To a solution of
4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)eth-
oxy)aniline XXIV (619 mg, 2.17 mmol) and
6,8-dibromoimidazo[1,2-a]pyrazine (601 mg, 2.2 mmol) in IPA (15 mL)
was added N,N-Diisopropylethylamine (0.95 ml, 5.43 mmol). The
mixture was stirred at 110.degree. C. for 16 h. After this time,
DCM (10 mL) and sat aqueous NaHCO.sub.3 (15 mL) were added. The
aqueous layer was separated and extracted with DCM (2.times.10 mL).
The combined organic extracts were washed with brine (10 mL) and
dried over sodium sulfate. The resulting residue was purified by
column chromatography ISCO Rf (24 g column) eluting with a gradient
of 100% DCM--60:35:5 DCM:Et.sub.2O:MeOH to provide
6-bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-
-yl)oxy)ethoxy)phenyl)imidazo[1,2-a]pyrazin-8-amine XXV.
tert-Butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-
-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)carbamate
XXVI
6-Bromo-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-
-yl)oxy)ethoxy)phenyl)imidazo[1,2-a]pyrazin-8-amine XXV (1.2 g, 2.4
mmol) was reacted according to the analogous method described in
Intermediate Example 1.01 (conversion of III to IV) to provide
tert-butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(-
2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)carbamate XXVI.
tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a-
]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyra-
n-2-yl)oxy)ethoxy)phenyl)carbamate XXVII
tert-Butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(-
2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)carbamate XXVI was
reacted with XV according to the methods of CHEMISTRY B as
described in Example 2 to provide the desired compound tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy-
)ethoxy)phenyl)carbamate XXVII.
2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2-(4-(oxet-
an-3-yl)piperazin-1-yl)phenoxy)ethanol (5)
The compound tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy-
)ethoxy)phenyl)carbamate XXVII (313 mg, 0.35 mmol) was de-protected
by the analogous method described in Example 2 to provide
2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2-(4-(oxe-
tan-3-yl)piperazin-1-yl)phenoxy)ethanol (5). LCMS-ESI.sup.+ (m/z):
[M+H].sup.+: 504.3. .sup.1H NMR (300 MHz, d.sub.6-DMSO) .delta.:
9.52 (s, 1H), 8.61 (s, 1H), 8.51 (s, 1H), 8.14 (d, J=1.1 Hz, 1H),
7.89 (s, 1H), 7.81 (d, J=2.3 Hz, 1H), 7.74-7.60 (m, 2H), 6.90 (d,
J=8.6 Hz, 1H), 6.47 (s, 2H), 5.74 (s, 1H), 4.86-4.76 (m, 1H), 4.50
(dt, J=25.6, 6.3 Hz, 4H), 4.04 (t, J=5.1 Hz, 2H), 3.73 (q, J=5.1
Hz, 2H), 3.51-3.42 (m, 1H), 3.02 (s, 4H), 2.40 (s, 4H).
Example 6. Preparation of
2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)phenyl-
)piperazin-1-yl)methyl)propane-1,3-diol (6)
##STR00147##
Oxetane-3-carbaldehyde XXVIII
To a round-bottomed flask equipped with a stirring bar,
oxetan-3-ylmethanol (2.00 g, 22.7 mmol) was dissolved in DCM (50
mL) and Dess-Martin periodinane (10.67 g, 28.38 mmol) was added in
one portion. The reaction mixture was stirred at RT overnight. The
solids were filtered through celite, and washed with DCM (3
mL.times.5). The filtrate was removed and concentrated in vacuo and
the resulting crude oxetane-3-carbaldehyde XXVIII was used in the
next step directly.
1-(4-Nitrophenyl)-4-(oxetan-3-ylmethyl)piperazine XXIX
To a round-bottomed flask equipped with a stirring bar,
oxetane-3-carbaldehyde XXVIII (0.977 g, 11.35 mmol),
1-(4-nitrophenyl)piperazine (1.18 g, 5.68 mmol) in DCM (100 mL),
and HOAc (1.70 g, 28.38 mmol) in DCM (2 mL) were added. After 5
minutes, NaBH(OAc).sub.3 (24.06 g, 113.05 mmol) was added. The
resulting mixture was stirred at room temperature for 2 h. Most
volatiles were removed in vacuo. DCM (200 mL) was added, followed
by saturated NaHCO.sub.3 aqueous solution (20 mL), and the
resulting mixture was stirred for 20 minutes. The organic phase was
separated and washed with saturated NaHCO.sub.3 aqueous solution
(20 mL.times.3), brine (20 mL.times.1), dried over
Na.sub.2SO.sub.4, filtered and solvents were removed in vacuo. The
residue was passed through a silica gel column (MeOH:DCM=0:100 to
5:95 to 25:75) to provide the desired compound XXIX.
4-(4-(Oxetan-3-ylmethyl)piperazin-1-yl)aniline XXX
To a round-bottomed flask equipped with a stirring bar, were added
1-(4-nitrophenyl)-4-(oxetan-3-ylmethyl)piperazine XXIX (3.20 g,
11.54 mmol), ethanol (60 mL) and water (60 mL). Following the
addition of iron (4.51 g, 80.77 mmol) and ammonium chloride (4.32
g, 80.77 mmol), the reaction mixture was heated at 80.degree. C.
for 1 h, then filtered through Celite and washed with DCM (5
mL.times.5). The resulting filtrate was extracted with DCM (20
mL.times.3). The combined organic extracts were washed with water
(20 mL.times.2), brine (20 mL.times.1), dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The desired
4-(4-(oxetan-3-ylmethyl)piperazin-1-yl)aniline XXX was
obtained.
6-Bromo-N-(4-(4-(oxetan-3-ylmethyl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyr-
azin-8-amine XXXI
To a seal tube equipped with a stirring bar,
4-(4-(oxetan-3-ylmethyl)piperazin-1-yl)aniline XXX (1.19 g, 4.81
mmol), 6,8-dibromoimidazo[1,2-a]pyrazine (1.33 g, 4.81 mmol),
isopropanol (24.1 mL), and diisopropylethylamine (1.37 g, 10.58
mmol) were added, and the reaction mixture was heated at
100.degree. C. overnight. Most solvents were removed in vacuo and
DCM (200 mL) was added to the mixture. The solution was washed with
H.sub.2O (20 mL.times.2), brine (20 mL.times.1), dried over
Na.sub.2SO.sub.4, filtered and solvents were removed in vacuo. The
resulting residue was passed through a silica gel column
(MeOH:DCM=5:95) and light red solids were obtained as the desired
compound XXXI, 0.692 g.
tert-Butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(1-(oxetan-3-ylmethyl)pip-
eridin-4-yl)phenyl)carbamate XXXII
To a round-bottomed flask equipped with a stirring bar, were added
6-bromo-N-(4-(4-(oxetan-3-ylmethyl)piperazin-1-yl)phenyl)imidazo[1,2-a]py-
razin-8-amine XXXI (560 mg, 1.27 mmol), DCM (11 mL), di-tert-butyl
dicarbonate (414.4 mg, 1.90 mmol), and triethylamine (640.5 mg,
6.33 mmol). The reaction mixture was heated at 50.degree. C.
overnight. DCM (200 mL) was added, and the resulting solution was
washed with water (20 mL.times.2), brine (20 mL.times.1), dried
over Na.sub.2SO.sub.4, filtered and solvents were removed in vacuo.
Column chromatography gave the desired compound XXXII as yellow
solids.
tert-Butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a-
]pyrazin-8-yl)(4-(4-(oxetan-3-ylmethyl)piperazin-1-yl)phenyl)carbamate
XXXIII
To a round-bottomed flask equipped with a stirring bar, tert-butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-ylmethyl)piperazin-1-y-
l)phenyl)carbamate XXXII (150 mg, 0.276 mmol), N, N-BisBoc
6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazin-2-amine XV
(255.8 mg, 0.607 mmol) in DME (2.3 mL), Pd(PPh.sub.3).sub.4 (16.0
mg, 0.14 mmol), Na.sub.2CO.sub.3 aqueous solution (1.0 N, 0.91 mL,
0.91 mmol), and DME (2 mL) were added. The mixture was heated at
75.degree. C. for 2, then DCM (200 mL) was added and the resulting
mixture was washed with water (30 mL.times.3), brine (30
mL.times.1), dried over MgSO.sub.4, filtered, and solvents were
removed in vacuo. Purification by silica gel column (MeOH:DCM=5:95)
gave the desired compound XXXIII.
2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)phenyl)-
piperazin-1-yl)methyl)propane-1,3-diol (6)
To a solution of tert-butyl
(6-(6-(bis(tert-butoxycarbonyl)amino)pyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
-yl)(4-(4-(oxetan-3-ylmethyl)piperazin-1-yl)phenyl)carbamate XXXIII
(250 mg, 0.33 mmol) in DCM (30 mL) was added TFA (940.3 mg, 8.25
mmol). The resulting mixture was stirred at room temperature for
overnight. More TFA (752.2 mg, 6.60 mmol) was added and stirred at
room temperature overnight. Most solvents were removed in vacuo,
DCM (200 mL) and saturated NaHCO.sub.3 aqueous solution (30 mL)
were added and the resulting mixture was stirred for 30 minutes.
The organic phase was separated, washed with saturated NaHCO.sub.3
aqueous solution (20 mL.times.4), brine (20 mL.times.1). The
aqueous phase was extracted with DCM (30 mL.times.2). The combined
organic phases were washed with brine (20 mL.times.1), dried over
Na.sub.2SO.sub.4, filtered, and solvents were removed in vacuo. The
crude material was purified on ISCO column, MeOH:DCM=0:100 to 5:95
to 7.5:92.5 to 25:75 to elute the desired compounds. Two compounds
were obtained, the first is the oxetane compound; and the other the
desired compound 6. LCMS-ESI.sup.+ (m/z): [M+H].sup.+: 476. .sup.1H
NMR (300 MHz, d.sub.6-DMSO) .delta.: 9.51 (s, 1H), 8.60 (s, 1H),
8.49 (s, 1H), 8.14 (d, J=1.5 Hz, 1H), 7.95 (d, J=9 Hz, 2H), 7.90
(s, 1H), 7.64 (s, 1H), 6.99 (d, J=9 Hz, 2H), 6.48 (s, 2H), 4.51
(broad S, 2H), 3.43 (d, J=6 Hz, 4H), 3.12 (broad m, 4H), 2.54
(broad m, 4H), 2.34 (d, J=7.2 Hz, 2H), 1.83 (m, 1H).
Example 7. Preparation of
2-(5-((6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)--
2-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol (7)
##STR00148##
tert-butyl
tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3--
yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)amino-
)imidazo[1,2-a]pyrazin-6-yl)-3-chloropyrazin-2-yl)carbamate
XXXIV
A flask equipped with a reflux condenser was charged with
tert-butyl
(6-bromoimidazo[1,2-a]pyrazin-8-yl)(4-(4-(oxetan-3-yl)piperazin-1-yl)-3-(-
2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)carbamate XXVI
(prepared as described in Example 5) (352 mg, 0.52 mmol),
2-(bis-boc-amino)-3-chloro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl-
)pyrazine (prepared by analogous method as used in Example 2 for
the preparation of compound XV) (500 mg, 1.1 mmol),
Pd(PPh.sub.3).sub.4 (30 mg, 0.03 mmol) in sodium carbonate (1.6 mL,
1M in H.sub.2O) and DME (4.8 mL). The mixture was heated to reflux
for 1 h. The reaction was cooled to room temperature, diluted with
DCM and H.sub.2O. The aqueous layer was separated and extracted
with DCM. The combined organic extracts were washed with brine,
dried over sodium sulfate, filtered and concentrated under reduced
pressure. The resulting residue was purified by column
chromatography ISCO Rf (4 g column) eluting with a gradient of 100%
DCM--100% 60/35/5 DCM/Et.sub.2O/MeOH, appropriate fractions were
combined and concentrated to provide the desired compound
tert-butyl
tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperaz-
in-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)amino)imidazo[1-
,2-a]pyrazin-6-yl)-3-chloropyrazin-2-yl)carbamate XXXIV.
tert-butyl
tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3--
yl)piperazin-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)amino-
)imidazo[1,2-a]pyrazin-6-yl)-3-methylpyrazin-2-yl)carbamate
XXXV
A microwave vial was charged with tert-butyl
tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperaz-
in-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)amino)imidazo[1-
,2-a]pyrazin-6-yl)-3-chloropyrazin-2-yl)carbamate XXXIV (258 mg,
0.28 mmol), methylboronic acid (503 mg, 8.4 mmol),
Pd(PPh.sub.3).sub.4 (32 mg, 0.03 mmol) in sodium carbonate (0.8 mL,
1M in H.sub.2O) and DME (2.5 mL). The mixture was heated at
150.degree. C. for 20 min. The reaction was cooled to room
temperature, diluted with DCM and H.sub.2O. The aqueous layer was
separated and extracted with DCM. The combined organic extracts
were washed with brine, dried over sodium sulfate, filtered and
concentrated under reduced pressure. The resulting residue was
purified by column chromatography ISCO Rf (4 g column) eluting with
a gradient of 100% DCM--100% 75/18/7 DCM/Et.sub.2O/MeOH to provide
the desired compound tert-butyl
tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperaz-
in-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)amino)imidazo[1-
,2-a]pyrazin-6-yl)-3-methylpyrazin-2-yl)carbamate XXXV.
2-(5-((6-(6-Amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-2-
-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol (7)
To a solution of tert-butyl
tert-butoxycarbonyl(6-(8-((tert-butoxycarbonyl)(4-(4-(oxetan-3-yl)piperaz-
in-1-yl)-3-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)amino)imidazo[1-
,2-a]pyrazin-6-yl)-3-methylpyrazin-2-yl)carbamate XXXV (165 mg,
0.18 mmol) in DCM (2.2 mL) was added TFA (1.1 mL, 0.11 mmol). The
mixture was stirred at rt for 16 h. The reaction was diluted with
9:1 DCM:MeOH and H.sub.2O. The aqueous layer was separated and
extracted with 9:1 DCM:MeOH. The combined organic extracts were
washed with brine, dried over sodium sulfate, filtered and
concentrated under reduced pressure. The resulting residue was
purified by column chromatography eluting with a gradient of 100%
75/18/7 DCM/Et.sub.2O/MeOH--100% 70/20/10 DCM/Et.sub.2O/MeOH to
provide the desired compound
2-(5-((6-(6-amino-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-8-yl)amino)--
2-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol (7, 56 mg, 59%).
LCMS-ESI.sup.+ (m/z): [M+H].sup.+: 518.2. .sup.1H NMR (300 MHz,
d.sub.6-DMSO) .delta.: 9.49 (s, 1H), 8.56 (s, 1H), 8.44 (s, 1H),
8.13 (d, J=1.1 Hz, 1H), 7.85-7.66 (m, 2H), 7.62 (d, J=1.1 Hz, 1H),
6.90 (d, J=8.6 Hz, 1H), 6.25 (s, 2H), 4.87-4.77 (m, 1H), 4.50 (dt,
J=25.2, 6.3 Hz, 4H), 4.04 (t, J=5.1 Hz, 2H), 3.74 (q, J=5.2 Hz,
2H), 3.51-3.39 (m, 1H), 3.10-2.95 (m, 4H), 2.45-2.35 (m, 4H), 2.34
(s, 3H). Alternatively, compound XXXIV could be taken directly to
this step and similarly de-protected to provide the
5-chloropyrazine substituted analog.
Monomesylate and Succinate Forms
X-ray powder diffraction (XRPD) analysis of the monomesylate (MSA)
and succinate forms of the compound of Example 2 herein were
conducted on a diffractometer (PANanalytical XPERT-PRO, PANalytical
B.V., Almelo, Netherlands) using copper radiation (Cu K.alpha.,
.lamda.=1.5418 .ANG.). Samples were prepared for analysis by
depositing the powdered sample in the center of an aluminum holder
equipped with a zero background plate. The generator was operated
at a voltage of 45 kV and amperage of 40 mA. Slits used were Soller
0.02 rad., antiscatter 1.0.degree., and divergence. The sample
rotation speed was 2 sec. Scans were performed from 2 to 40.degree.
2-theta. Data analysis was performed by X'Pert Highscore version
2.2c (PANalytical B.V., Almelo, Netherlands) and X'Pert data viewer
version 1.2d (PANalytical B.V., Almelo, Netherlands). The XRPD
patterns for Mono MSA Forms I & II were obtained using the
instrument setting as follows: 45 KV, 40 mA, Cu K.alpha.,
.lamda.=1.5418 .ANG., scan range 2.-40.degree., step size
0.0167.degree., counting time: 15.875 s. The XRPD patterns for
Succinate Forms I & II were obtained using the instrument
setting as follows: 45 KV, 40 mA, Cu K.alpha., .lamda.=1.5418
.ANG., scan range 2.-40.degree., step size 0.0084.degree., counting
time: 95.250 s.
.sup.1H NMR spectra of the monomesylate (MSA) and succinate forms
of the compound of Example 2 were collected on a Varian 400-MR 400
MHz instrument with 7620AS sample changer. The default proton
parameters are as follows: spectral width: 14 to -2 ppm (6397.4
Hz); relaxation delay: 1 sec; acquisition time: 2.5559 sec; number
of scans or repetitions: 8; temperature: 25 C. Samples were
prepared in dimethyl sulfoxide-d.sub.6, unless otherwise stated.
Off-line analysis was carried out using MNova software.
Example
8--6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phe-
nyl)imidazo[1,2-a]pyrazin-8-amine monomesylate Form I
Methanesulfonic acid (MSA) salt Form I was prepared by dissolving
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine (Example 2) in 11 volumes of
acetone/H.sub.2O (36:64 vol. %) with 1 molar equivalent of methane
sulfonic acid (MSA) at room temperature. The solution was then
charged with 19 volumes of acetone over 1 hour and the reactor
contents were stirred at room temperature overnight.
XRPD analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form I was conducted as
described above and provided the diffraction pattern seen in FIG.
1, with the peaks in the table below.
TABLE-US-00007 Pos. Rel. Int. No. [.degree.2Th.] [%] 1 19.6606 100
2 17.2746 93.07 3 17.8971 69.96 4 21.6306 65.74 5 25.7805 59.16 6
18.7593 51.5 7 13.7252 48.77 8 15.7206 41.91 9 24.7364 38.09 10
18.4345 36.84
In one embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form I may be characterized by
XRPD peaks 19.7 (19.6606), 17.3 (17.2746), 17.9 (17.8971), 21.6
(21.6306), and 25.8 (25.7805). In a further embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form I may be characterized by
XRPD peaks 19.7 (19.6606), 17.3 (17.2746), 17.9 (17.8971), and 21.6
(21.6306). In another embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form I may be characterized by
XRPD peaks 6.0, 6.2, 8.6, and 9.6.
NMR Analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Mono MSA Salt Form I, conducted as
described above, provided the NMR spectrum seen in FIG. 2.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.57 (s, 1H), 9.60 (s,
1H), 8.62 (s, 1H), 8.47 (s, 1H), 8.17 (d, J=1.2 Hz, 1H), 8.03-7.96
(m, 2H), 7.90 (s, 1H), 7.69 (d, J=1.2 Hz, 1H), 7.09 (d, J=9.0 Hz,
2H), 4.78 (p, J=8.0 Hz, 4H), 4.49 (m, 1H), 4.00-2.8 (m, 10H), 2.32
(s, 3H).
Differential Scanning Calorimetry (DSC):
DSC was performed for each of the examples indicated herein using a
TA Instruments Q2000 DSC instrument. The sample was placed into an
aluminum DSC pan, and the weight accurately recorded. The pan was
covered with a lid, and then either crimped or hermetically sealed.
The same cell was heated under a nitrogen purge at a rate of
10.degree. C./min, up to a final temperature of 300.degree. C.
Indium was used as the calibration standard.
A DSC analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Mono MSA Salt Form I, conducted as
described above, is seen in FIG. 3.
Thermogravimetric Analysis (TGA):
TGA was performed for each of the examples indicated herein using a
TA Instruments Q5000 TGA instrument. Each sample was placed in an
aluminum sample pan and inserted into the TG furnace. The furnace
was heated under nitrogen at a rate of 10.degree. C./min, up to a
final temperature of 300.degree. C. The TGA furnace was calibrated
using the magnetic Curie point method.
A TGA analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Mono MSA Salt Form I, conducted as
described above, is seen in FIG. 4.
Example
9--6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phe-
nyl)imidazo[1,2-a]pyrazin-8-amine monomesylate Form II
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Mono MSA Salt Form II was prepared by
drying
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Mono MSA Salt Form I (Example 8) in a
vacuum oven at .about.40.degree. C. with a N.sub.2 purge.
XRPD analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form II was conducted as
described above and provided the diffraction pattern seen in FIG.
5, with the peaks in the table below.
TABLE-US-00008 Pos. Rel. Int. No. [.degree.2Th.] [%] 1 17.2698 100
2 25.1384 67.84 3 20.4423 63.66 4 19.5732 62.11 5 18.5264 50.36 6
17.7884 50.07 7 21.6273 45.52 8 15.2397 44 9 6.855 35.01 10 13.65
26
In one embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form II may be characterized
by XRPD peaks 17.3 (17.2698), 25.1 (25.1384), 20.4 (20.4423), 19.6
(19.5732), and 18.5 (18.5264). In an additional embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form II may be characterized
by XRPD peaks 17.3 (17.2698), 25.1 (25.1384), 20.4 (20.4423), and
19.6 (19.5732). In another embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine monomesylate Form II may be characterized
by XRPD peaks 6.1, 6.9, 11.0, and 13.6.
NMR Analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Mono MSA Salt Form II, conducted as
described above, provided the NMR spectrum seen in FIG. 6.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.61 (s, 1H), 8.63 (s,
1H), 8.47 (s, 1H), 8.19 (d, J=1.2 Hz, 1H), 8.02-7.95 (m, 2H), 7.91
(s, 1H), 7.72 (d, J=1.2 Hz, 1H), 7.13-7.06 (m, 2H), 4.85-4.72 (m,
4H), 4.53-4.45 (m, 1H), 4.30-2.75 (m, 10H), 2.34 (s, 3H).
A DSC analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Mono MSA Salt Form II, conducted as
described above, is seen in FIG. 7.
A TGA analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Mono MSA Salt Form II, conducted as
described above, is seen in FIG. 8.
Example
10--6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)ph-
enyl)imidazo[1,2-a]pyrazin-8-amine succinate Form I
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form I was prepared by first
dissolving 1.6 mol. eq. of succinic acid in THF, and then charging
the acidic solution to
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine. The material was then stirred at room
temperature with a magnetic stir bar overnight.
XRPD analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine succinate Form I was conducted as described
above and provided the diffraction pattern seen in FIG. 9, with the
peaks in the table below.
TABLE-US-00009 Pos. Rel. Int. No. [.degree.2Th.] [%] 1 16.5 100 2
24.5 38.64 3 17.7 9.27 4 28.4 8.68 5 21.8 7.57 6 8.0 6.53 7 23.1
4.59 8 12.1 4.38 9 8.3 3.78 10 27.1 3.65
In one embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form I may be characterized by
XRPD peaks 16.5, 24.5, 17.7, 28.4, and 21.8. In another embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form I may be characterized by
XRPD peaks 16.5, 24.5, 8.0 and 8.3.
NMR Analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form I, conducted as described
above, provided the NMR spectrum seen in FIG. 10.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.12 (s, 2H), 9.48 (s,
1H), 8.59 (s, 1H), 8.48 (s, 1H), 8.12 (d, J=1.1 Hz, 1H), 7.97-7.86
(m, 3H), 7.62 (d, J=1.1 Hz, 1H), 7.01-6.94 (m, 2H), 6.45 (s, 2H),
4.55 (t, J=6.5 Hz, 2H), 4.46 (t, J=6.1 Hz, 2H), 3.49-3.38 (m, 1H),
3.13 (t, J=4.9 Hz, 4H), 2.40 (s, 10H).
A DSC analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form I, conducted as described
above, is seen in FIG. 11.
A TGA analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form I, conducted as described
above, is seen in FIG. 12.
The process for preparing
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine succinate Form I was also repeated using
IPA, Acetone, and 2-MeTHF as solvents.
Example
11--6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)ph-
enyl)imidazo[1,2-a]pyrazin-8-amine succinate Form II
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine free base was charged with 10.0 parts
2-propanol, followed by rapid agitation, to form a slurry. A
separate solution of succinic acid (0.43 parts, 1.6 mol eq.) in
2-propanol (15 parts) was prepared at ambient temperature and was
added to the slurry. The resulting slurry was then agitated at
ambient temperature for about 1 day. Another solution of succinic
acid (0.09 parts, 0.3 mol eq.) in 2-propanol (3 parts) was added to
the slurry and the resulting slurry was agitated at ambient
temperature for about two days. An additional solution of succinic
acid (0.27 parts, 1.0 mol eq.) in 2-propanol (8 parts) was prepared
at ambient temperature and added to the slurry and the resulting
slurry was agitated for about 2 days. Then the content temperature
was adjusted to 40.degree. C. and the slurry was agitated for about
two hours. The content was then returned to ambient temperature and
agitated for about 16 hours. The resulting slurry was then
filtered, rinsed with 2-propanol (7.0 parts), and dried at
60.degree. C.
XRPD analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine succinate Form II was conducted as
described above and provided the diffraction pattern seen in FIG.
13, with the peaks in the table below.
TABLE-US-00010 Pos. Rel. Int. No. [.degree.2Th.] [%] 1 24.9821 100
2 16.3186 38.39 3 21.952 18.44 4 7.8958 17.62 5 7.5828 6.9 6
28.5998 6.52 7 11.3329 5.73 8 30.8568 5.48 9 28.0273 5.21 10
21.5026 4.73
In one embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form II may be characterized by
XRPD peaks 25.0 (24.9821), 16.3 (16.3186), 22.0 (21.952), 7.9
(7.8958), and 7.6 (7.5828). In a further embodiment
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form II may be characterized by
XRPD peaks 25.0 (24.9821), 16.3 (16.3186), 7.9 (7.8958), and 7.6
(7.5828).
NMR Analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form II, conducted as described
above, provided the NMR spectrum seen in FIG. 14.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.13 (s, 2H), 9.48 (s,
1H), 8.58 (s, 1H), 8.47 (s, 1H), 8.12 (d, J=1.1 Hz, 1H), 7.97-7.86
(m, 3H), 7.62 (d, J=1.1 Hz, 1H), 7.02-6.94 (m, 2H), 6.45 (s, 2H),
4.55 (t, J=6.5 Hz, 2H), 4.46 (t, J=6.0 Hz, 2H), 3.44 (p, J=6.3 Hz,
1H), 3.17-3.10 (m, 4H), 2.40 (s, 10H), 1.02 (d, J=6.1 Hz, 2H).
A DSC analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form II, conducted as described
above, is seen in FIG. 15.
A TGA analysis of
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-yl)phenyl)imidaz-
o[1,2-a]pyrazin-8-amine Succinate Form II, conducted as described
above, is seen in FIG. 16.
Biological Examples
Example 12: High Throughput Syk Biochemical Assay
Syk activity was measured using KinEASE (Cisbio), a time-resolved
fluorescence resonance energy transfer (TR-FRET) immunoassay. In
this assay, Syk-catalyzes the phosporylation of a XL665-labeled
peptide substrate. Europium conjugated phospho-tyrosine specific
antibody binds the resulting phosphorylated peptide. Formation of
phosphorylated peptide is quantified by TR-FRET with Europium as
the donor and XL665 the acceptor in a 2-step endpoint assay. In
brief, test compounds serially diluted in DMSO were delivered into
Corning white, low volume, non-binding 384 well plates using the
Echo 550 acoustic liquid dispenser (Labcyte.RTM.). Syk enzyme and
substrates were dispensed into assay plates using a Multi-Flo
(Bio-Tek Instruments). The standard 5 .mu.L reaction mixture
contained 20 .mu.M ATP, 1 .mu.M biotinylated peptide, 0.015 nM of
Syk in reaction buffer (50 mM Hepes, pH 7.0, 0.02% NaN.sub.3, 0.1%
BSA, 0.1 mM Orthovanadate, 5 mM MgCl.sub.2, 1 mM DTT, 0.025%
NP-40). After 30 minutes of incubation at room temperature, 5 .mu.L
of Stop and Detect Solution (1:200 Europium Cryptate labeled
anti-phosphorylated peptide antibody solution and 125 nM
strepavidin-XL665 Tracer in a 50 mM Hepes pH 7.0 detection buffer
containing sufficient EDTA) was added. The plate was then further
incubated for 120 minutes at room temperature and read using an
Envision 2103 Multilabeled reader (PerkinElmer) with
excitation/emission/FRET emission at 340 nm/615 nm/665 nm,
respectively. Fluorescence intensities at 615 nm and 665 nm
emission wavelengths were expressed as a ratio (665 nm/615 nm).
Percent inhibition was calculated as follows: 10%
Inhibition=100.times.(Ratio.sub.Sample-Ratio.sub.0%
Inhibition)/(Ratio.sub.100% Inhibition-Ratio.sub.0% Inhibition)
where 0.1% DMSO (0% inhibition) was the negative control and 1
.mu.M K252a (100% inhibition) was used as the positive control.
Activity of the compounds of Examples 1-7 are provided in the
following table, demonstrating the compounds are Syk inhibitors
with IC.sub.50 below 50 nM.
TABLE-US-00011 Syk IC.sub.50 Example No.: Compound Name (nM) Ex.-1:
6-(6-amino-5-methylpyrazin-2-yl)-N-(4-(4-(oxetan-3- 6.2
yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-2:
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1- 13.5
yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-3:
(R)-(4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8- 13.3
yl)amino)phenyl)morpholin-2-yl)methanol Ex.-4:
6-(6-aminopyrazin-2-yl)-5-methyl-N-(4-(4-(oxetan-3- 44
yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-5:
2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8- 12.2
yl)amino)-2-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol Ex.-6:
2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8- 14.5
yl)amino)phenyl)piperazin-1-yl)methyl)propane-1,3-diol Ex.-7:
2-(5-((6-(6-ami- 8.7 no-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-
8-yl)amino)-2-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol
Example 13: 384-Well HTBS Whole Blood CD63 Basophil Assay
Syk activity was assessed in relation to reduced activation of
basophils as measured by the expression of CD63 in a human whole
blood basophil cellular assay (25% blood). Basophil activation was
measured in human whole blood using the Flow CAST kit (Buhlmann
Laboratories AG, Baselstrasse, Switzerland) following the protocol
provided by the manufacturer with minor modifications. Fresh human
whole blood in heparin was collected and delivered same day
(AllCells, Emeryville, Calif.). Whole blood samples were incubated
with either DMSO (1% final) or serial diluted compounds in DMSO for
60 minutes at 37.degree. C. Basophils were activated using the
anti-FceRI mAb and stained with anti-CD63-FITC and anti-CCR3-PE for
20 minutes at 37.degree. C. (per well: 50 .mu.L of whole blood was
mixed with 113 .mu.L of stimulation buffer, 8.5 .mu.L anti-FceRI
mAb, 8.5 .mu.L Ab stain CCR3-PE/CD63-FITC). Cells were centrifuged
at 1000.times.g for 18 minutes and 150 .mu.L/well of supernatant
removed. Red blood cells were lysed and cells fixed by 2 rounds of
cell lysing: resuspending cell pellets with 150 .mu.L/well 1.times.
lysis buffer, incubating at room temperature for 10 minutes, and
collecting cell pellets by centrifuging for 1200 rpms for 5
minutes. Cells were washed with 150 L/well wash buffer twice, and
resuspended in a final volume of 75 L/well of wash buffer for
either immediate flow cytometery analysis or overnight incubation
at 4.degree. C. followed by flow cytometry analysis. Degranulation
(basophil activation) was detected by CD63 surface expression on
CCR3 positive cells. The percent CD63 positive cells within the
gated basophil population were determined and normalized to the
DMSO (negative control) and control compound (positive control).
Activity of the compounds of Examples 1-7 are provided in the
following table, demonstrating the compounds are effective in
reducing the activation of basophils, with EC.sub.50 below 200
nM.
TABLE-US-00012 CD63 EC.sub.50 Example No.: Compound Name (nM)
Ex.-1: 6-(6-amino-5-methylpyrazin-2-yl)-N-(4-(4-(oxetan-3- 51
yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-2:
6-(6-aminopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1- 80
yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-3:
(R)-(4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8- 63
yl)amino)phenyl)morpholin-2-yl)methanol Ex.-4:
6-(6-aminopyrazin-2-yl)-5-methyl-N-(4-(4-(oxetan-3- 157
yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-5:
2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8- 120
yl)amino)-2-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol Ex.-6:
2-((4-(4-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8- 128
yl)amino)phenyl)piperazin-1-yl)methyl)propane-1,3-diol Ex.-7:
2-(5-((6-(6-ami- 167 no-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-
8-yl)amino)-2-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol
Example 14: Kinetic Solubility
The kinetic solubility of compounds in phosphate buffer at pH 7.4
was assessed. The compounds to be tested were dissolved in
dimethylsulfoxide at a 10 mM concentration. Stock samples were
diluted, 3 .mu.l with 297 .mu.l of the phosphate buffer at pH 7.4
(DulBecco's phosphate buffered saline (Sigma-Aldrich D8662),
overall molarity is 0.149M and pH 7.43). The samples were then
incubated for 24 hours at 37.degree. C. with shaking, the
centrifuged and an aliquot taken and tested relative to a known
standard concentration of 0.1 mM. The kinetic solubility of the
compounds of Examples 1-7 are provided in the following table,
demonstrating the compounds have kinetic solubility at pH 7.4 of
greater than 90 .mu.M.
TABLE-US-00013 Solubility pH 7.4 Example No.: Compound Name (.mu.M)
Ex.-1: 6-(6-amino-5-methylpyrazin-2-yl)-N-(4-(4-(oxetan-3- 95
yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-2:
6-(6-ami- 95 nopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-
yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-3:
(R)-(4-(4-((6-(6-ami- 91 nopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
yl)amino)phenyl)morpholin-2-yl)methanol Ex.-4:
6-(6-aminopyrazin-2-yl)-5-methyl-N-(4-(4-(oxetan-3- 100
yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-5:
2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8- 97
yl)amino)-2-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol Ex.-6:
2-((4-(4-((6-(6-ami- 99 nopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
yl)amino)phenyl)piperazin-1-yl)methyl)propane-1,3-diol
Example 15: Human Hepatocyte Stability Assay
The human hematocyte stability of the compounds as predicted
hepatocyte clearance in L/hr/kg was assessed. Compounds to be
tested were diluted to 200 .mu.M (4 .mu.l of 10 mM DMSO stock into
196 .mu.l ACN:H.sub.2O (50:50). Propranolol was used as a positive
control, and buffer only without hepatocytes as 0% control. These
were further diluted 4 .mu.l with 891 .mu.l KHB buffer (InVitroGRO
catalog number Z99074) to provide 2.times. dosing solution. To each
well of 24 well plate, 250 .mu.l of the 2.times. dosing solution
was added to each well with 250 .mu.l of hepatocytes cells
(1.times.10.sup.6 viable cells/ml per well) or KHB for control
samples to achieve a final compound concentration of 1 .mu.M during
incubation. The final solvent concentration was 0.01% DMSO and
0.25% ACN. The culture plate was placed on a rocker and incubated
at 37.degree. C., 5% CO.sub.2. Samples were collected at time 0, 1,
3, and 6 hours. The loss of parent compound was determined using
LC-MS methods against a standard curve. Activity of the compounds
of Examples 1-7 are provided in the following table, showing
hepatocyte clearance of about 0.12 L/hr/kg or less.
TABLE-US-00014 Hheps CL Example No.: Compound Name (L/hr/kg) Ex.-1:
6-(6-amino-5-methylpyrazin-2-yl)-N-(4-(4-(oxetan-3- 0.12
yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-2:
6-(6-ami- 0.055 nopyrazin-2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-1-
yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-3:
(R)-(4-(4-((6-(6-ami- 0.09 nopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
yl)amino)phenyl)morpholin-2-yl)methanol Ex.-4:
6-(6-aminopyrazin-2-yl)-5-methyl-N-(4-(4-(oxetan-3- 0.08
yl)piperazin-1-yl)phenyl)imidazo[1,2-a]pyrazin-8-amine Ex.-5:
2-(5-((6-(6-aminopyrazin-2-yl)imidazo[1,2-a]pyrazin-8- 0.07
yl)amino)-2-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol Ex.-6:
2-((4-(4-((6-(6-ami- 0.08 nopyrazin-2-yl)imidazo[1,2-a]pyrazin-8-
yl)amino)phenyl)piperazin-1-yl)methyl)propane-1,3-diol Ex.-7:
2-(5-((6-(6-ami- 0.05
no-5-methylpyrazin-2-yl)imidazo[1,2-a]pyrazin-
8-yl)amino)-2-(4-(oxetan-3-yl)piperazin-1-yl)phenoxy)ethanol
Example 16: Comparison to Known Syk Inhibitors
The assays of Examples 8-11 were used to compare the compounds as
described herein with compounds known in the art. The data
comparing the compounds of Examples 1-7 to previously described
compounds is provided in the following table. From these results,
it is clear that compounds as described herein are desirable as Syk
inhibitors, with improved Syk and CD63 activity relative to the
known compounds, improved kinetic solubility (at least about 9-fold
more soluble) and hepatocyte clearance (at least about 2-fold less
clearance). As such, the combination of improved Syk and CD63
inhibitory activity with improved kinetic solubility and clearance
provides compounds that are expected to be effective at treating
diseases as described herein with improved pharmacokinetic
properties.
TABLE-US-00015 Syk CD63 Solubility Hheps IC.sub.50 IC.sub.50 pH 7.4
CL Compound Name (nM) (nM) (.mu.M) (units) Ex.-1:
6-(6-amino-5-methylpyrazin- 6.2 51 95 0.12
2-yl)-N-(4-(4-(oxetan-3-yl)piperazin-
1-yl)phenyl)imidazo[1,2-a]pyrazin-8- amine Ex.-2:
6-(6-aminopyrazin-2-yl)-N- 13.5 80 95 0.055
(4-(4-(oxetan-3-yl)piperazin-1- yl)phenyl)imidazo[1,2-a]pyrazin-8-
amine Ex.-3: (R)-(4-(4-((6-(6- 13.3 63 91 0.09
aminopyrazin-2-yl)imidazo[1,2- a]pyrazin-8-
yl)amino)phenyl)morpholin-2- yl)methanol Ex.-4:
6-(6-aminopyrazin-2-yl)-5- 44 157 100 0.08
methyl-N-(4-(4-(oxetan-3- yl)piperazin-1-
yl)phenyl)imidazo[1,2-a]pyrazin-8- amine Ex.-5:
2-(5-((6-(6-aminopyrazin-2- 12.2 120 97 0.07
yl)imidazo[1,2-a]pyrazin-8- yl)amino)-2-(4-(oxetan-3-
yl)piperazin-1-yl)phenoxy)ethanol Ex.-6:
2-((4-(4-((6-(6-aminopyrazin- 14.5 128 99 0.08
2-yl)imidazo[1,2-a]pyrazin-8- yl)amino)phenyl)piperazin-1-
yl)methyl)propane-1,3-diol Ex.-7: 2-(5-((6-(6-amino-5- 8.7 167 nd
0.05 methylpyrazin-2-yl)imidazo[1,2-
a]pyrazin-8-yl)amino)-2-(4-(oxetan-
3-yl)piperazin-1-yl)phenoxy)ethanol Known compounds:
6-(5-aminopyridin-3-yl)-N-(4- 31 101 5 0.68
morpholinophenyl)imidazo[1,2- a]pyrazin-8-amine
6-(3-aminophenyl)-N-(3,4- 188 809 3 0.24
dimethoxyphenyl)imidazo[1,2- a]pyrazin-8-amine
6-(5-amino-6-methylpyridin-3-yl)-N- 16 250 5 0.80
(4-morpholinophenyl)imidazo[1,2- a]pyrazin-8-amine
6-(6-aminopyridin-3-yl)-N-(3,4- 53 734 10 0.90
dimethoxyphenyl)imidazo[1,2- a]pyrazin-8-amine
Throughout this specification, various patents, patent applications
and other types of publications (e.g., journal articles) are
referenced. The disclosure of all patents, patent applications, and
publications cited herein are hereby incorporated by reference in
their entirety for all purposes.
* * * * *
References